CN101637188A - Method for preventing and curing fungal diseases of plants - Google Patents
Method for preventing and curing fungal diseases of plants Download PDFInfo
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- CN101637188A CN101637188A CN 200810224679 CN200810224679A CN101637188A CN 101637188 A CN101637188 A CN 101637188A CN 200810224679 CN200810224679 CN 200810224679 CN 200810224679 A CN200810224679 A CN 200810224679A CN 101637188 A CN101637188 A CN 101637188A
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Abstract
The invention discloses a preparation method of a microbial inoculum for preventing and curing fungal diseases of plants; the microbial inoculum for preventing and curing fungal diseases of plants isprepared by mixing Trichoderma longibrachiatum and Trichoderma citrinoviride. The microbial inoculum prepared by the method of the invention is used to carry out control efficiency experiments and theresults show that the control efficiency of liquid microbial inoculum to cucumber blight is up to 54.8% and the control efficiency to green pepper phytophthora disease is up to 67.9%; the control efficiency of solid microbial inoculum to cucumber blight is up to 45.12% and the yield of cucumber is increased by 11.06% so that the effect for preventing diseases and increasing yield is obvious; thecontrol efficiency of solid microbial inoculum to green pepper phytophthora disease is up to 61.90% and the average plant height of the green pepper in experimental group is increased by 8.92% compared with that of the green pepper in control group and the yield of green pepper is increased by 37.19% so that the microbial inoculum of the invention is shown to have obvious effect for preventing increases, promoting the growth and increasing the yield.
Description
Technical field
The present invention relates to a kind of method of preventing and treating fungal diseases of plants.
Background technology
Since the eighties in 20th century, China's growing vegetables already develops rapidly, and vegetables have become the second largest crops that are only second to cereal crops.Because the particular surroundings of vegetable growing, suitable condition is provided for generation, the development of vegetable disease, cause kind, quantity and the extent of injury of vegetable disease all to present the trend that increases year by year, have a strong impact on output and the quality of vegetables, cause very big economic loss to the vegetable grower, become " bottleneck " that restriction China growing vegetables already develops and strengthen competitiveness in the international market.The control of vegetable disease, be China's growing vegetables already develop essential, also be vegetable grower's an urgent demand.
Vegetables mostly are year after year continuous cropping plantation, thereby cause the pathogen of some disease to accumulate in a large number, and especially some soil-borne diseases are on the rise, and increase the weight of year by year as the harm of cucumber fusarium axysporum, eggplant verticillium wilt and green pepper eqpidemic disease etc.For a long time, aspect vegetable disease prevented and treated, the agricultural chemicals particularly use of chemical pesticide had been brought into play enormous function.Because characteristics such as chemical pesticide have instant effect, fungicidal spectrum is wide and easy to use, development becomes one of effective means of diseases prevention high yield rapidly in disease control.But for a long time, use chemical pesticide can cause beneficial microbe to be killed or to suppress repeatedly and in a large number, destroy biology community structure, cause environmental pollution, make in the vegetables harmful substance residual in a large number, cause pathogen to produce drug resistance or pesticide resistance, thereby the consumption of chemical pesticide is increased, brought more serious problem.The sustainable development that the present age is agriculture requires the producer when utilizing resource, improving output, notes protection and improves the environment that people depend on for existence, and this has just proposed stern challenge to traditional concept and bad way in the vegetable disease control.Owing to the long-term chemical agent that uses is prevented and treated reasons such as environmental destruction that various vegetable disease causes and the enhancing of pathogen pesticide resistance, press for and seek a kind of safer and more effective vegetable disease controlling way.The characteristics of biological control have just in time remedied the deficiency of chemical control, thereby become the research direction that receives much attention.
Utilize biological method control vegetable disease not only to avoid chemical pesticide to bring a series of problems, and safety, effectively and lasting, and be difficult for producing drug resistance,, noresidue nontoxic, free from environmental pollution, can keep ecological balance and the better simply characteristics of production technology the person poultry safety.Because biological control has the ecological and disease-resistant double effects of protection concurrently, in the vegetable disease control, bring into play enormous function, thereby obtaining countries in the world researcher's concern, oneself is very important and obtain the effective measures paid attention to day by day in preventing and treating through becoming vegetable disease.
At present, found that multiple microorganism has the effect of biological and ecological methods to prevent plant disease, pests, and erosion, Trichoderma is exactly a class ubiquity and have the biological and ecological methods to prevent plant disease, pests, and erosion beneficial bacterium of important economic implications wherein.(Biological control agents BCAs) is subjected to common concern to Trichoderma (Trichoderma spp.) always as a kind of important disease flocking biocontrol factor.External existing commercial wooden removing mildew comes out, as the Topshield (Trichoderma harzianum T22) of the U.S. and the Trichodex (Trichoderma harzianum T39) of Israel.
Cucumber fusarium axysporum and green pepper eqpidemic disease are the most common and endanger two serious class silborne fungal diseases in the growing vegetables, and it is wilting and wilt the vascular bundle obstruction of root and stem, browning etc. that cardinal symptom shows as plant.In recent years, become one of main disease of vegetables by the fusarium wilt due to the sharp sickle spore bacterium (Fusarium oxysporum Schlecht.), wherein, it is the most serious to be injured with cucumber, the incidence of disease is generally 20%～30%, and the heavy regional incidence of disease of being injured is up to 80%～90%, even total crop failure.The green pepper eqpidemic disease is a kind of soil-borne disease that is caused by green pepper epidemic disease mould (Phytophthora capsici Leonian), is the destructive disease of green pepper, all can fall ill in the whole growing of green pepper.Broad irrigation and hot and humid condition cause that easily the large tracts of land of green pepper eqpidemic disease is popular, often cause the green pepper underproduction, even total crop failure, and the green pepper eqpidemic disease has become the destructive disease of green pepper in the world wide.Effective control of cucumber fusarium axysporum and green pepper eqpidemic disease becomes a difficult problem anxious to be solved in the vegetables production.
Summary of the invention
The purpose of this invention is to provide a kind of method of preventing and treating fungal diseases of plants.
The microbial inoculum of control fungal diseases of plants provided by the present invention, its active component comprise long handle wood mould (Trichoderma longibrachiatum) and lemon green trichoderma (Trichoderma citrinoviride).
Mould and the colony forming unit number ratio lemon green trichoderma of described long handle wood is 1: (0.8-1.2), be specially 1: 1.
Described long handle wood mould (Trichoderma longibrachiatum) specifically can be long handle wood mould (Trichodermalongibrachiatum) ACCC 30150, and described lemon green trichoderma (Trichoderma citrinoviride) specifically can be lemon green trichoderma (Trichoderma citrinoviride) ACCC 30152.
The active ingredient that is used to prevent and treat the microbial inoculum of fungal diseases of plants provided by the present invention also can only be made up of and lemon green trichoderma mould by described long handle wood.
In the above-mentioned microbial inoculum, the mould and described lemon green trichoderma of described long handle wood can be distinguished independent packaging, also may be mixed together.
The microbial inoculum of control fungal diseases of plants of the present invention can be liquid bacterial agent, also can be solid fungicide, adds adsorbing base and can obtain solid fungicide in liquid bacterial agent; In the described solid fungicide, described adsorbing base is the mixture of being made up of chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite.
The ratio of weight and number of described chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite is 1: (0.38-0.42): (0.18-0.22): (0.18-0.22): (0.18-0.22), be specially 1: 0.4: 0.2: 0.2: 0.2.
In the above-mentioned solid fungicide, the proportioning of described active component and described adsorbing base is 4.5 * 10
8The CFU spore: (1.6-1.7) g adsorbing base is specially 4.5 * 10
8CFU spore: 1.65g adsorbing base.
Described fungal disease is by at least a caused fungal disease in following 11 kinds of fungies: cucumber rhizoctonia rot bacterium (R.solani), cucumber fusarium axysporum (F.oxysporum), dosporium cucumerinumand its (C.cucumerinum), tomato early blight bacterium (A.solani), botrytis cinerea (B.cinerea), eggplant verticillium wilt bacterium (V.dahliae), Chinese cabbage alternaria (A.brassicae), fusarium graminearum (F.graminearum), root rotof flax bacterium (B.sorokinianum), spot defoliation bacterium (A.mali), green pepper eqpidemic disease germ (P.capsici).
The present invention utilizes the microbial inoculum of above-mentioned preparation to carry out control efficiency experiment, and the result shows that liquid bacterial agent reaches 54.8% to the control efficiency of cucumber fusarium axysporum, and the control efficiency of green pepper eqpidemic disease is reached 67.9%.Solid fungicide reaches 45.12% to the control efficiency of cucumber fusarium axysporum, and makes cucumber production promoting 11.06%, its diseases prevention and obvious effect of increasing production; Solid fungicide reaches 61.90% to the control efficiency of green pepper eqpidemic disease, and the average plant height of experimental group green pepper increases by 8.92% than control group, green pepper volume increase 37.19%, illustrate microbial inoculum of the present invention diseases prevention, short give birth to and the effect of volume increase remarkable.
Embodiment
The used bacterial classification of following examples all comes from INST OF AGRICULTURAL RESOURCES Chinese agriculture microorganism fungus kind preservation administrative center and (is called for short ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, INST OF AGRICULTURAL RESOURCES, postcode 100081).
The preparation of the microbial inoculum of embodiment 1, control fungal diseases of plants and control efficiency experiment thereof
1, the preparation of the liquid bacterial agent of control fungal diseases of plants
Chlamydospore medium: KH is produced in screening
2PO
42.0g/L, NH
4NO
31.0g/L, FeSO
4.7H
2O 0.005g/L, MnSO
40.0016g/L, CaCl
20.3g/L, NaCl 1.0g/L, MgSO
4.7H
2O 0.3g/L, corn stalk powder (crossing the 40mm sieve) 10g/L.PH?6.0。Sterilization is 20 minutes under 115 ℃ of conditions.
Long handle mould ACCC 30150 of wood and lemon green trichoderma ACCC 30152 are inoculated into respectively in the PDA liquid nutrient medium, and 30 ℃ of shaking tables are cultivated 3 days (265 rev/mins of shaking speed).Be inoculated into above-mentioned screening according to the inoculum concentration of 10% (volumn concentration) then and produce in the chlamydospore medium, 30 ℃ of shaking tables cultivate that chlamydosporic amount reaches 4.5 * 10 in the zymotic fluid of (250 rev/mins of shaking speed) and lemon green trichoderma mould to long handle wood
8CFU/ml, the zymotic fluid that above-mentioned long handle wood is mould and the zymotic fluid of lemon green trichoderma mix according to 1: 1 volume ratio, obtain preventing and treating the liquid bacterial agent of fungal diseases of plants.
2, the preparation of the solid fungicide of control fungal diseases of plants
With chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite according to 5: 2: 1: 1: 1 mass ratio mixes, and obtains preventing and treating the adsorbing base of the solid fungicide of fungal diseases of plants.With 1.65g adsorbing base and contain 4.5 * 10
8The liquid bacterial agent that the above-mentioned steps 1 of CFU spore obtains mixes mutually, obtains preventing and treating the solid fungicide of fungal diseases of plants.
3, the control efficiency of microbial inoculum experiment
(1) liquid bacterial agent control cucumber fusarium axysporum test
With temperature is seed 12～24h of 37 ℃ sterile water immersion cucumber (variety name: Chang Chun Mi Ci is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the seedling-cultivating tray, behind seed sprouting,, cover skim soil more in the above by row sowing, water spray is preserved moisture, and when treating cucumber seedling length to 2～3 slice true leaf, carries out cucumber fusarium axysporum control test.
It is 10 that cucumber fusarium axysporum germ (ACCC 30442) is made spore concentration
6The bacteria suspension of individual/mL is completely cured above-mentioned cucumber seedling earlier, soaks 5min again in the bacteria suspension of above-mentioned cucumber fusarium axysporum germ, transplants engagement then, and every seedling inoculates the bacteria suspension 5mL of above-mentioned cucumber fusarium axysporum germ, covers skim soil.Then, with the liquid bacterial agent of the control fungal diseases of plants of the cucumber seedling of above-mentioned inoculation cucumber fusarium axysporum germ inoculation above-mentioned steps 1 preparation, every seedling inoculation 2.5mL, earthing is cultivated.Simultaneously with the cucumber of the normal growth of not inoculating the cucumber fusarium axysporum germ as negative control (CKo), the cucumber of liquid bacterial agent of control fungal diseases of plants that does not inoculate above-mentioned steps 1 preparation with inoculation cucumber fusarium axysporum germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is as shown in table 1 to the control efficiency of cucumber fusarium axysporum.
Table 1 liquid bacterial agent is to the control efficiency of cucumber fusarium axysporum
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 54.8% to the control efficiency of cucumber fusarium axysporum.
(2) solid fungicide control cucumber fusarium axysporum test
Cucumber seedling was transplanted in 1 heart stage of leaf, 4 days 3 May, was benchmark with the traditional cultivation management of milpa, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Three processing are established in test altogether, be respectively: normal fertilising group (CK1 group), (every seedling is except normal fertilising for normal fertilising+matrix group (CK2 group), use 10g matrix again, spread manuer in holes under cucumber root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for the solid fungicide group, use the solid fungicide of 10g above-mentioned steps 2 again, spread manuer in holes under cucumber root).4 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.2 row cucumber are transplanted in every sub-district, and line-spacing is 65cm, and spacing in the rows is 29cm, amounts to 12 sub-districts.Duration of test every investigation in 20 days once writes down diseased plant number and dead strain number, calculates diseased plant rate and dead plant rate.Three repetitions are established in experiment.The field incidence result of different disposal group cucumber fusarium axysporum is as shown in table 2.
The field incidence of table 2 different disposal group cucumber fusarium axysporum
The result shows, under same management condition, compares with the CK1 group, and the diseased plant rate of CK2 group and solid fungicide group and dead plant rate all have decline in various degree.Wherein after the field planting 15 days the time, three experimental group all do not have dead strain.The CK2 group is organized decline 18.25%-22.22% with the diseased plant rate of solid fungicide group than CK1; Afterwards, the difference of diseased plant rate and dead plant rate is more and more littler between CK1 group and the CK2 group, and diseased plant rate and dead plant rate difference between solid fungicide group and CK1 group and the CK2 group are increasing.The solid fungicide that shows above-mentioned steps 2 preparations has the better prevention effect to cucumber fusarium axysporum.
The initial phase of morbidity and the control efficiency of different disposal group cucumber fusarium axysporum are as shown in table 3.
Table 3 solid fungicide is to the control efficiency of cucumber fusarium axysporum
The result shows, solid fungicide group and CK2 group were than the disease time of CK1 group all late 5 days, and its protection effect you can well imagine high 45.12% and 4.04% than the CK1 component, and the solid fungicide group is extremely remarkable with the protection effect difference that CK1 organizes and CK2 organizes.
Above-mentioned gradation results cucumber, the calculating output of weighing respectively respectively handled.The result is as shown in table 4.
Table 4 solid fungicide is to the effect of increasing production of cucumber
The result shows that the solid fungicide group is than CK1 group cucumber production promoting 11.06%, significant difference.
Above-mentioned experimental result shows, the solid fungicide diseases prevention and the obvious effect of increasing production of above-mentioned steps 2 preparations behind the solid fungicide of inoculation above-mentioned steps 2, can improve the resistance against diseases of cucumber, alleviate the loss that the cucumber fusarium axysporum evil causes.
(3) liquid bacterial agent control green pepper eqpidemic disease test
With temperature is the seed 24h of 37 ℃ sterile water immersion green pepper (variety name: eggplant door green pepper is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the dish for cultivating, behind seed sprouting, directly sowing is covered skim soil in the above again in cultivation tray, and water spray is preserved moisture, and when the green pepper seedling grows to 5～6 true leaves, carries out the prevention and control of sweetbell redpepper epidemic disease test.
It is 10 that green pepper eqpidemic disease germ (ACCC 30093) is made spore concentration
3The bacteria suspension of individual/mL is not transplanted the bacteria suspension that direct filling root is inoculated above-mentioned green pepper eqpidemic disease germ with above-mentioned green pepper seedling, and the bacteria suspension of every the above-mentioned green pepper eqpidemic disease of seedling inoculation 5mL germ covers skim soil.Then, the green pepper seedling of above-mentioned inoculation green pepper eqpidemic disease germ is inoculated the liquid bacterial agent of the control fungal diseases of plants of above-mentioned steps 1 preparation, every seedling inoculation 3mL, earthing is cultivated.Simultaneously with the green pepper of not inoculating green pepper eqpidemic disease germ normal growth as negative control (CKo), the green pepper of liquid bacterial agent of control fungal diseases of plants of not inoculating above-mentioned steps 1 preparation with inoculation green pepper eqpidemic disease germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is as shown in table 5 to the control efficiency of green pepper eqpidemic disease.
Table 5 liquid bacterial agent is to the control efficiency of green pepper eqpidemic disease
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 67.9% to the control efficiency of green pepper eqpidemic disease.
(4) solid fungicide control green pepper eqpidemic disease test
The green pepper seedling was transplanted in 4 leaf phases, and managing with the traditional cultivation of milpa is benchmark, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Five processing are established in test altogether, be respectively: normal fertilising group (CKo group), (every seedling is except normal fertilising for normal fertilising+low dosage matrix group (CK1 group), use 10g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for normal fertilising+high dose matrix group (CK2 group), use 20g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is used the solid fungicide of 10g above-mentioned steps 2 to low dosage solid fungicide group again except normal fertilising, spread manuer in holes under the green pepper root), high dose solid fungicide group (every seedling is used the solid fungicide of 20g above-mentioned steps 2 again except normal fertilising, spread manuer in holes under the green pepper root).3 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.Every ridge plants 2 row green peppers, row spacing 50cm, and per two ridges are a sub-district (19.2m
2), totally 15 sub-districts.Planting density 3300 strains/667m
2Duration of test every investigation in 20 days once writes down diseased plant number and state of an illness rank, calculates diseased plant rate and disease index.Three repetitions are established in experiment.The field incidence and the control efficiency of different disposal group green pepper eqpidemic disease are as shown in table 6.
Table 6 solid fungicide is to the control efficiency of green pepper eqpidemic disease
The result shows that in the solid fungicide of above-mentioned steps 2 preparations, high dose solid fungicide group is 61.90% to the control efficiency of green pepper eqpidemic disease, and low dosage solid fungicide group is 45.95% to the control efficiency of green pepper eqpidemic disease.
The different disposal group to the green pepper plant height to influence the result as shown in table 7.
Table 7 solid fungicide is to the influence of green pepper plant height
The result shows that the solid fungicide of above-mentioned steps 2 preparations has certain facilitation to the growth of green pepper.The average plant height of low dosage microbial inoculum group green pepper increases by 4.34% than the CKo group, and the average plant height of high dose microbial inoculum group green pepper increases by 8.92% than the CKo group.
Above-mentioned gradation results green pepper, the calculating output of weighing respectively respectively handled.The result is as shown in table 8.
Table 8 solid fungicide is to the effect of increasing production of green pepper
The result shows that high dose microbial inoculum group is than CKo group green pepper volume increase 37.19%, significant difference.
Above-mentioned experimental result shows, the solid fungicide of above-mentioned steps 2 preparations, its diseases prevention and obvious effect of increasing production, behind the solid fungicide of inoculation above-mentioned steps 2, the plant height of green pepper obviously improves, but also can improve the resistance against diseases of green pepper, alleviates the loss that green pepper eqpidemic disease disease causes.
The preparation of the microbial inoculum of embodiment 2, control fungal diseases of plants and control efficiency experiment thereof
1, the preparation of the liquid bacterial agent of control fungal diseases of plants
Chlamydospore medium: KH is produced in screening
2PO
42.0g/L, NH
4NO
31.0g/L, FeSO
4.7H
2O 0.005g/L, MnSO
40.0016g/L, CaCl
20.3g/L, NaCl 1.0g/L, MgSO
4.7H
2O 0.3g/L, corn stalk powder (crossing the 40mm sieve) 10g/L.PH?6.0。Sterilization is 20 minutes under 115 ℃ of conditions.
Long handle mould ACCC 30150 of wood and lemon green trichoderma ACCC 30152 are inoculated into respectively in the PDA liquid nutrient medium, and 30 ℃ of shaking tables are cultivated 3 days (265 rev/mins of shaking speed).Be inoculated into above-mentioned screening according to the inoculum concentration of 10% (volumn concentration) then and produce in the chlamydospore medium, 30 ℃ of shaking tables cultivate that chlamydosporic amount all reaches 4.5 * 10 in the zymotic fluid of (250 rev/mins of shaking speed) and lemon green trichoderma mould to long handle wood
8CFU/ml, the zymotic fluid that above-mentioned long handle wood is mould and the zymotic fluid of lemon green trichoderma mix according to 1: 1.2 volume ratio, obtain preventing and treating the liquid bacterial agent of fungal diseases of plants.
2, the preparation of the solid fungicide of control fungal diseases of plants
With chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite according to 1: 0.42: 0.22: 0.22: 0.22 mass ratio mixes, and obtains preventing and treating the adsorbing base of the solid fungicide of fungal diseases of plants.With 1.7g adsorbing base and contain 4.5 * 10
8The liquid bacterial agent that the above-mentioned steps 1 of CFU spore obtains mixes mutually, obtains preventing and treating the solid fungicide of fungal diseases of plants.
3, the control efficiency of microbial inoculum experiment, concrete process of the test is with embodiment 1.
(1) liquid bacterial agent control cucumber fusarium axysporum test
With temperature is seed 12～24h of 37 ℃ sterile water immersion cucumber (variety name: Chang Chun Mi Ci is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the seedling-cultivating tray, behind seed sprouting,, cover skim soil more in the above by row sowing, water spray is preserved moisture, and when treating cucumber seedling length to 2～3 slice true leaf, carries out cucumber fusarium axysporum control test.
It is 10 that cucumber fusarium axysporum germ (ACCC 30442) is made spore concentration
6The bacteria suspension of individual/mL is completely cured above-mentioned cucumber seedling earlier, soaks 5min again in the bacteria suspension of above-mentioned cucumber fusarium axysporum germ, transplants engagement then, and every seedling inoculates the bacteria suspension 5mL of above-mentioned cucumber fusarium axysporum germ, covers skim soil.Then, with the liquid bacterial agent of the control fungal diseases of plants of the cucumber seedling of above-mentioned inoculation cucumber fusarium axysporum germ inoculation above-mentioned steps 1 preparation, every seedling inoculation 2.5mL, earthing is cultivated.Simultaneously with the cucumber of the normal growth of not inoculating the cucumber fusarium axysporum germ as negative control (CKo), the cucumber of liquid bacterial agent of control fungal diseases of plants that does not inoculate above-mentioned steps 1 preparation with inoculation cucumber fusarium axysporum germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is as shown in table 9 to the control efficiency of cucumber fusarium axysporum.
Table 9 liquid bacterial agent is to the control efficiency of cucumber fusarium axysporum
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 51.8% to the control efficiency of cucumber fusarium axysporum.
(2) solid fungicide control cucumber fusarium axysporum test
Cucumber seedling was transplanted in 1 heart stage of leaf, 4 days 3 May, was benchmark with the traditional cultivation management of milpa, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Three processing are established in test altogether, be respectively: normal fertilising group (CK1 group), (every seedling is except normal fertilising for normal fertilising+matrix group (CK2 group), use 10g matrix again, spread manuer in holes under cucumber root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for the solid fungicide group, use the solid fungicide of 10g above-mentioned steps 2 again, spread manuer in holes under cucumber root).4 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.2 row cucumber are transplanted in every sub-district, and line-spacing is 65cm, and spacing in the rows is 29cm, amounts to 12 sub-districts.Duration of test every investigation in 20 days once writes down diseased plant number and dead strain number, calculates diseased plant rate and dead plant rate.Three repetitions are established in experiment.The initial phase of morbidity and the control efficiency of different disposal group cucumber fusarium axysporum are as shown in table 10.
Table 10 solid fungicide is to the control efficiency of cucumber fusarium axysporum
The result shows, solid fungicide group and CK2 group were than the disease time of CK1 group all late 5 days, and its protection effect you can well imagine high 45.04% and 3.98% than the CK1 component, and the solid fungicide group is extremely remarkable with the protection effect difference that CK1 organizes and CK2 organizes.
Above-mentioned gradation results cucumber, the calculating output of weighing respectively respectively handled.The result is as shown in table 11.
Table 11 solid fungicide is to the effect of increasing production of cucumber
The result shows that the solid fungicide group is than CK1 group cucumber production promoting 10.90%, significant difference.
Above-mentioned experimental result shows, the solid fungicide diseases prevention and the obvious effect of increasing production of above-mentioned steps 2 preparations behind the solid fungicide of inoculation above-mentioned steps 2, can improve the resistance against diseases of cucumber, alleviate the loss that the cucumber fusarium axysporum evil causes.
(3) liquid bacterial agent control green pepper eqpidemic disease test
With temperature is the seed 24h of 37 ℃ sterile water immersion green pepper (variety name: eggplant door green pepper is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the dish for cultivating, behind seed sprouting, directly sowing is covered skim soil in the above again in cultivation tray, and water spray is preserved moisture, and when the green pepper seedling grows to 5～6 true leaves, carries out the prevention and control of sweetbell redpepper epidemic disease test.
It is 10 that green pepper eqpidemic disease germ (ACCC 30093) is made spore concentration
3The bacteria suspension of individual/mL is not transplanted the bacteria suspension that direct filling root is inoculated above-mentioned green pepper eqpidemic disease germ with above-mentioned green pepper seedling, and the bacteria suspension of every the above-mentioned green pepper eqpidemic disease of seedling inoculation 5mL germ covers skim soil.Then, the green pepper seedling of above-mentioned inoculation green pepper eqpidemic disease germ is inoculated the liquid bacterial agent of the control fungal diseases of plants of above-mentioned steps 1 preparation, every seedling inoculation 3mL, earthing is cultivated.Simultaneously with the green pepper of not inoculating green pepper eqpidemic disease germ normal growth as negative control (CKo), the green pepper of liquid bacterial agent of control fungal diseases of plants of not inoculating above-mentioned steps 1 preparation with inoculation green pepper eqpidemic disease germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is as shown in table 12 to the control efficiency of green pepper eqpidemic disease.
Table 12 liquid bacterial agent is to the control efficiency of green pepper eqpidemic disease
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 65.3% to the control efficiency of green pepper eqpidemic disease.
(4) solid fungicide control green pepper eqpidemic disease test
The green pepper seedling was transplanted in 4 leaf phases, and managing with the traditional cultivation of milpa is benchmark, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Five processing are established in test altogether, be respectively: normal fertilising group (CKo group), (every seedling is except normal fertilising for normal fertilising+low dosage matrix group (CK1 group), use 10g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for normal fertilising+high dose matrix group (CK2 group), use 20g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is used the solid fungicide of 10g above-mentioned steps 2 to low dosage solid fungicide group again except normal fertilising, spread manuer in holes under the green pepper root), high dose solid fungicide group (every seedling is used the solid fungicide of 20g above-mentioned steps 2 again except normal fertilising, spread manuer in holes under the green pepper root).3 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.Every ridge plants 2 row green peppers, row spacing 50cm, and per two ridges are a sub-district (19.2m
2), totally 15 sub-districts.Planting density 3300 strains/667m
2Duration of test every investigation in 20 days once writes down diseased plant number and state of an illness rank, calculates diseased plant rate and disease index.Three repetitions are established in experiment.The field incidence result of different disposal group green pepper eqpidemic disease is as shown in table 13.
Table 13 solid fungicide is to the control efficiency of green pepper eqpidemic disease
The result shows that in the solid fungicide of above-mentioned steps 2 preparations, high dose solid fungicide group is 59.48% to the control efficiency of green pepper eqpidemic disease, and low dosage solid fungicide group is 45.08% to the control efficiency of green pepper eqpidemic disease.
The different disposal group to the green pepper plant height to influence the result as shown in table 14.
Table 14 solid fungicide is to the influence of green pepper plant height
The result shows that the solid fungicide of above-mentioned steps 2 preparations has certain facilitation to the growth of green pepper.The average plant height of low dosage microbial inoculum group green pepper increases by 4.04% than the CKo group, and the average plant height of high dose microbial inoculum group green pepper increases by 8.62% than the CKo group.
Above-mentioned gradation results green pepper, the calculating output of weighing respectively respectively handled.The result is as shown in Table 15.
Table 15 solid fungicide is to the effect of increasing production of green pepper
The result shows that high dose microbial inoculum group is than CKo group green pepper volume increase 35.56%, significant difference.
Above-mentioned experimental result shows, the solid fungicide of above-mentioned steps 2 preparations, its diseases prevention and obvious effect of increasing production, behind the solid fungicide of inoculation above-mentioned steps 2, the plant height of green pepper obviously improves, but also can improve the resistance against diseases of green pepper, alleviates the loss that green pepper eqpidemic disease disease causes.
The preparation of the microbial inoculum of embodiment 3, control fungal diseases of plants and control efficiency experiment thereof
1, the preparation of the liquid bacterial agent of control fungal diseases of plants
Chlamydospore medium: KH is produced in screening
2PO
42.0g/L, NH
4NO
31.0g/L, FeSO
4.7H
2O 0.005g/L, MnSO
40.0016g/L, CaCl
20.3g/L, NaCl 1.0g/L, MgSO
4.7H
2O 0.3g/L, corn stalk powder (crossing the 40mm sieve) 10g/L.PH?6.0。Sterilization is 20 minutes under 115 ℃ of conditions.
Long handle mould ACCC 30150 of wood and lemon green trichoderma ACCC 30152 are inoculated into respectively in the PDA liquid nutrient medium, and 30 ℃ of shaking tables are cultivated 3 days (265 rev/mins of shaking speed).Be inoculated into above-mentioned screening according to the inoculum concentration of 10% (volumn concentration) then and produce in the chlamydospore medium, 30 ℃ of shaking tables cultivate that chlamydosporic amount all reaches 4.5 * 10 in the zymotic fluid of (250 rev/mins of shaking speed) and lemon green trichoderma mould to long handle wood
8CFU/ml, the zymotic fluid that above-mentioned long handle wood is mould and the zymotic fluid of lemon green trichoderma mix according to 1: 0.8 volume ratio, obtain preventing and treating the liquid bacterial agent of fungal diseases of plants.
2, the preparation of the solid fungicide of control fungal diseases of plants
With chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite according to 1: 0.38: 0.18: 0.18: 0.18 mass ratio mixes, and obtains preventing and treating the adsorbing base of the solid fungicide of fungal diseases of plants.With 1.6g adsorbing base and contain 4.5 * 10
8The liquid bacterial agent that the above-mentioned steps 1 of CFU spore obtains mixes mutually, obtains preventing and treating the solid fungicide of fungal diseases of plants.
3, the control efficiency of microbial inoculum experiment, concrete process of the test is with embodiment 1.
(1) liquid bacterial agent control cucumber fusarium axysporum test
With temperature is seed 12～24h of 37 ℃ sterile water immersion cucumber (variety name: Chang Chun Mi Ci is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the seedling-cultivating tray, behind seed sprouting,, cover skim soil more in the above by row sowing, water spray is preserved moisture, and when treating cucumber seedling length to 2～3 slice true leaf, carries out cucumber fusarium axysporum control test.
It is 10 that cucumber fusarium axysporum germ (ACCC 30442) is made spore concentration
6The bacteria suspension of individual/mL is completely cured above-mentioned cucumber seedling earlier, soaks 5min again in the bacteria suspension of above-mentioned cucumber fusarium axysporum germ, transplants engagement then, and every seedling inoculates the bacteria suspension 5mL of above-mentioned cucumber fusarium axysporum germ, covers skim soil.Then, with the liquid bacterial agent of the control fungal diseases of plants of the cucumber seedling of above-mentioned inoculation cucumber fusarium axysporum germ inoculation above-mentioned steps 1 preparation, every seedling inoculation 2.5mL, earthing is cultivated.Simultaneously with the cucumber of the normal growth of not inoculating the cucumber fusarium axysporum germ as negative control (CKo), the cucumber of liquid bacterial agent of control fungal diseases of plants that does not inoculate above-mentioned steps 1 preparation with inoculation cucumber fusarium axysporum germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is shown in table 16 to the control efficiency of cucumber fusarium axysporum.
Table 16 liquid bacterial agent is to the control efficiency of cucumber fusarium axysporum
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 52.7% to the control efficiency of cucumber fusarium axysporum.
(2) solid fungicide control cucumber fusarium axysporum test
Cucumber seedling was transplanted in 1 heart stage of leaf, 4 days 3 May, was benchmark with the traditional cultivation management of milpa, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Three processing are established in test altogether, be respectively: normal fertilising group (CK1 group), (every seedling is except normal fertilising for normal fertilising+matrix group (CK2 group), use 10g matrix again, spread manuer in holes under cucumber root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for the solid fungicide group, use the solid fungicide of 10g above-mentioned steps 2 again, spread manuer in holes under cucumber root).4 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.2 row cucumber are transplanted in every sub-district, and line-spacing is 65cm, and spacing in the rows is 29cm, amounts to 12 sub-districts.Duration of test every investigation in 20 days once writes down diseased plant number and dead strain number, calculates diseased plant rate and dead plant rate.Three repetitions are established in experiment.Three repetitions are established in experiment.The initial phase of morbidity and the control efficiency of different disposal group cucumber fusarium axysporum are shown in table 17.
Table 17 solid fungicide is to the control efficiency of cucumber fusarium axysporum
The result shows, solid fungicide group and CK2 group were than the disease time of CK1 group all late 5 days, and its protection effect you can well imagine high 45.08% and 4.01% than the CK1 component, and the solid fungicide group is extremely remarkable with the protection effect difference that CK1 organizes and CK2 organizes.
Above-mentioned gradation results cucumber, the calculating output of weighing respectively respectively handled.The result is shown in table 18.
Table 18 solid fungicide is to the effect of increasing production of cucumber
The result shows that the solid fungicide group is than CK1 group cucumber production promoting 10.98%, significant difference.
Above-mentioned experimental result shows, the solid fungicide diseases prevention and the obvious effect of increasing production of above-mentioned steps 2 preparations behind the solid fungicide of inoculation above-mentioned steps 2, can improve the resistance against diseases of cucumber, alleviate the loss that the cucumber fusarium axysporum evil causes.
(3) liquid bacterial agent control green pepper eqpidemic disease test
With temperature is the seed 24h of 37 ℃ sterile water immersion green pepper (variety name: eggplant door green pepper is provided by Chinese Academy of Agricultural Sciences vegetables and flowers research institute), vernalization.With the cultivating soil mixing, be contained in the dish for cultivating, behind seed sprouting, directly sowing is covered skim soil in the above again in cultivation tray, and water spray is preserved moisture, and when the green pepper seedling grows to 5～6 true leaves, carries out the prevention and control of sweetbell redpepper epidemic disease test.
It is 10 that green pepper eqpidemic disease germ (ACCC 30093) is made spore concentration
3The bacteria suspension of individual/mL is not transplanted the bacteria suspension that direct filling root is inoculated above-mentioned green pepper eqpidemic disease germ with above-mentioned green pepper seedling, and the bacteria suspension of every the above-mentioned green pepper eqpidemic disease of seedling inoculation 5mL germ covers skim soil.Then, the green pepper seedling of above-mentioned inoculation green pepper eqpidemic disease germ is inoculated the liquid bacterial agent of the control fungal diseases of plants of above-mentioned steps 1 preparation, every seedling inoculation 3mL, earthing is cultivated.Simultaneously with the green pepper of not inoculating green pepper eqpidemic disease germ normal growth as negative control (CKo), the green pepper of liquid bacterial agent of control fungal diseases of plants of not inoculating above-mentioned steps 1 preparation with inoculation green pepper eqpidemic disease germ is as positive control (CK).The cultivation of preserving moisture, all " Invest, Then Investigate " result of the tests.
Three repetitions are established in experiment, and the liquid bacterial agent of above-mentioned steps 1 preparation is shown in table 19 to the control efficiency of green pepper eqpidemic disease.
Table 19 liquid bacterial agent is to the control efficiency of green pepper eqpidemic disease
The result shows that the liquid bacterial agent of above-mentioned steps 1 preparation can reach 65.7% to the control efficiency of green pepper eqpidemic disease.
(4) solid fungicide control green pepper eqpidemic disease test
The green pepper seedling was transplanted in 4 leaf phases, and managing with the traditional cultivation of milpa is benchmark, and moisture and other control measures are consistent.Chemical agent prevention and control disease is not all used in all experiment sub-districts.Five processing are established in test altogether, be respectively: normal fertilising group (CKo group), (every seedling is except normal fertilising for normal fertilising+low dosage matrix group (CK1 group), use 10g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is except normal fertilising for normal fertilising+high dose matrix group (CK2 group), use 20g matrix again, spread manuer in holes under the green pepper root, matrix is the adsorbing base of above-mentioned steps 2 preparation solid fungicides), (every seedling is used the solid fungicide of 10g above-mentioned steps 2 to low dosage solid fungicide group again except normal fertilising, spread manuer in holes under the green pepper root), high dose solid fungicide group (every seedling is used the solid fungicide of 20g above-mentioned steps 2 again except normal fertilising, spread manuer in holes under the green pepper root).3 repetitions are established in every kind of processing, and each tests the sub-district randomized arrangement.Every ridge plants 2 row green peppers, row spacing 50cm, and per two ridges are a sub-district (19.2m2), totally 15 sub-districts.Planting density 3300 strains/667m
2Duration of test every investigation in 20 days once writes down diseased plant number and state of an illness rank, calculates diseased plant rate and disease index.Three repetitions are established in experiment.The field incidence result of different disposal group green pepper eqpidemic disease is shown in table 20.
Table 20 solid fungicide is to the control efficiency of green pepper eqpidemic disease
The result shows that in the solid fungicide of above-mentioned steps 2 preparations, high dose solid fungicide group is 59.87% to the control efficiency of green pepper eqpidemic disease, and low dosage solid fungicide group is 45.63% to the control efficiency of green pepper eqpidemic disease.
The different disposal group to the green pepper plant height to influence the result shown in table 21.
Table 21 solid fungicide is to the influence of green pepper plant height
The result shows that the solid fungicide of above-mentioned steps 2 preparations has certain facilitation to the growth of green pepper.The average plant height of low dosage microbial inoculum group green pepper increases by 4.18% than the CKo group, and the average plant height of high dose microbial inoculum group green pepper increases by 8.75% than the CKo group.
Above-mentioned gradation results green pepper, the calculating output of weighing respectively respectively handled.The result is shown in table 22.
Table 22 solid fungicide is to the effect of increasing production of green pepper
The result shows that high dose microbial inoculum group is than CK1 group green pepper volume increase 36.21%, significant difference.
Above-mentioned experimental result shows, the solid fungicide of above-mentioned steps 2 preparations, its diseases prevention and obvious effect of increasing production, behind the solid fungicide of inoculation above-mentioned steps 2, the plant height of green pepper obviously improves, but also can improve the resistance against diseases of green pepper, alleviates the loss that green pepper eqpidemic disease disease causes.
Claims (10)
1, a kind of method of preventing and treating fungal diseases of plants is with the microbial inoculum of control fungal diseases of plants fungal diseases of plants to be prevented and treated;
The active component of the microbial inoculum of described control fungal diseases of plants comprises long handle wood mould (Trichodermalongibrachiatum) and lemon green trichoderma (Trichoderma citrinoviride).
2, method according to claim 1 is characterized in that: the mould and colony forming unit number ratio lemon green trichoderma of described long handle wood is 1: (0.8-1.2).
3, microbial inoculum according to claim 2 is characterized in that: the mould and colony forming unit number ratio lemon green trichoderma of described long handle wood is 1: 1.
4, according to arbitrary described method among the claim 1-3, it is characterized in that: described long handle wood mould (Trichoderma longibrachiatum) is long handle wood mould (Trichoderma longibrachiatum) ACCC30150, and described lemon green trichoderma (Trichoderma citrinoviride) is lemon green trichoderma (Trichodermacitrinoviride) ACCC 30152.
5, according to arbitrary described method among the claim 1-4, it is characterized in that: comprise adsorbing base in the described microbial inoculum, described adsorbing base is the mixture of being made up of chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite.
6, method according to claim 5, it is characterized in that: in the described adsorbing base, the ratio of weight and number of described chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite is 1: (0.38-0.42): (0.18-0.22): (0.18-0.22): (0.18-0.22).
7, method according to claim 6 is characterized in that: the ratio of weight and number of described chicken manure, wheat bran, soybean cake powder, the peat composed of rotten mosses and vermiculite is for being 1: 0.4: 0.2: 0.2: 0.2.
8, according to arbitrary described method among the claim 1-7, it is characterized in that: the proportioning of described active component and described adsorbing base is 4.5 * 10
8CFU spore: (1.6-1.7) g adsorbing base.
9, method according to claim 8 is characterized in that: the proportioning of described active component and described adsorbing base is 4.5 * 10
8CFU spore: 1.65g adsorbing base.
10, according to arbitrary described method among the claim 1-9, it is characterized in that: described fungal disease is by at least a caused fungal disease in following 11 kinds of fungies: cucumber rhizoctonia rot bacterium (R.solani), cucumber fusarium axysporum (F.oxysporum), dosporium cucumerinumand its (C.cucumerinum), tomato early blight bacterium (A.solani), botrytis cinerea (B.cinerea), eggplant verticillium wilt bacterium (V.dahliae), Chinese cabbage alternaria (A.brassicae), fusarium graminearum (F.graminearum), root rotof flax bacterium (B.sorokinianum), spot defoliation bacterium (A.mali) and green pepper eqpidemic disease germ (P.capsici).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102010825A (en) * | 2011-01-20 | 2011-04-13 | 上海创博生态工程有限公司 | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof |
| CN102212485A (en) * | 2011-05-09 | 2011-10-12 | 中国农业科学院农业资源与农业区划研究所 | Trichoderma longibrachiatum and application thereof to preventing and treating vegetable diseases |
| CN102523957A (en) * | 2011-12-14 | 2012-07-04 | 天津市山旮旯红果种植专业合作社 | Method for controlling peach tree scab |
| CN104920479A (en) * | 2015-07-10 | 2015-09-23 | 上海万力华生物科技有限公司 | Herbicide and preparation method and application thereof |
| US11812751B1 (en) | 2022-10-03 | 2023-11-14 | King Faisal University | Biocontrol agents for use against soil and air-borne fungal pathogens |
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2008
- 2008-10-23 CN CN 200810224679 patent/CN101637188A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102010825A (en) * | 2011-01-20 | 2011-04-13 | 上海创博生态工程有限公司 | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof |
| CN102010825B (en) * | 2011-01-20 | 2013-05-29 | 上海创博生态工程有限公司 | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof |
| CN102212485A (en) * | 2011-05-09 | 2011-10-12 | 中国农业科学院农业资源与农业区划研究所 | Trichoderma longibrachiatum and application thereof to preventing and treating vegetable diseases |
| CN102212485B (en) * | 2011-05-09 | 2012-12-19 | 中国农业科学院农业资源与农业区划研究所 | Trichoderma longibrachiatum and application thereof in preventing and treating vegetable diseases |
| CN102523957A (en) * | 2011-12-14 | 2012-07-04 | 天津市山旮旯红果种植专业合作社 | Method for controlling peach tree scab |
| CN104920479A (en) * | 2015-07-10 | 2015-09-23 | 上海万力华生物科技有限公司 | Herbicide and preparation method and application thereof |
| CN104920479B (en) * | 2015-07-10 | 2018-07-10 | 上海万力华生物科技有限公司 | A kind of herbicide and its preparation method and application |
| US11812751B1 (en) | 2022-10-03 | 2023-11-14 | King Faisal University | Biocontrol agents for use against soil and air-borne fungal pathogens |
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