CN110771631B - Method for preventing and treating nematode diseases by using composite nematode-killing microorganisms - Google Patents
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Abstract
The invention relates to a method for preventing and treating nematode diseases by using a compound nematicidal microorganism, which is prepared from gliocladium roseum (Gliocladium roseum)Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyceslilacinus(Thom.) Samson), Burkholderia arabica (B.arabic)Burkholderia arboris) Serratia zoophila (A), (B), (CSarratia entomophilus) Four microorganisms; the Burkholderia arabidopsis isBurkholderia arborisJ211; the preservation number is CCTCC NO: m2019747. 1) The burkholderia arabidopsis is a strain obtained by new research, and the strain has a high prevention and treatment effect on nematodes, and a new function is in continuous research; 2) on the basis of preventing and treating the nematodes by the burkholderia arabica, the other three strains are mixed, and the related volume ratio is controlled, so that the results of remarkable prevention and treatment effect and high prevention and treatment rate are realized; 3) the compound nematicidal microorganism has the capability of obviously improving the nematicidal capability of field crops; meanwhile, the commodity quality of the crops can be improved, and the yield of the crops can be improved.
Description
Technical Field
The invention belongs to the technical field of biological control; in particular to the technical field of a method for preventing and controlling plant nematode diseases by utilizing compound nematode-killing microorganisms.
Background
Nematodes are a very common plant pest in the growth process of crops. Some plant parasitic nematodes destroy the root tissue of plants, resulting in stunted root development and impaired nutrient absorption. Some nematodes also spread other diseases. Nematodes are estimated to cause losses equivalent to $ 1000 billion per year to crops.
The common nematode preventing and controlling method mainly comprises fumigation and chemical pesticide killing, and the green method is to use microbial inoculum with nematode killing function. The fumigation has a great destructive effect on the environment, and the use of the fumigation is forbidden by the state. The chemically synthesized pesticide has various hazards due to residues, harm to human bodies, environmental damage and ecological balance damage. Although nematode diseases of some plants can be controlled by traditional rotation, soil improvement and the like, rotation disease prevention is difficult to implement or even impossible to implement for some economic crops and forest trees. Biological control is more and more concerned by people because of the advantages of removing harm and increasing yield, protecting ecological balance, reducing environmental pollution and the like. The microorganism has the advantages of nature, no residue and the like, and becomes the method with the greatest development prospect. However, the existing biological control methods focus on using a specific single bacterial strain to control nematodes, and the single microbial inoculum has limited nematode killing effect, thereby influencing the popularization and use effects of biological agents. The invention improves the prevention and control effect of the microorganism for killing the nematodes by compounding the nematode killing microorganisms with different functions.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the existing biocontrol preparation for the field crop nematode disease has the problem of unsatisfactory control effect. Provides a method for preventing and controlling plant nematode diseases by using compound nematode-killing microorganisms.
The invention is realized by adopting the following technical scheme.
The invention relates to a method for preventing and controlling plant nematode diseases by using composite nematocidal microbe, which is prepared from gliocladium roseum (Gliocladium roseum)Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyceslilacinus(Thom.) Samson), Burkholderia arabica (B.arabic)Burkholderia arboris) Serratia zoophila (A), (B), (CSarratia entomophilus) Four microorganisms.
Further, the Burkholderia arabinonii of the invention isBurkholderia arborisJ211; the preservation number is CCTCC NO: m2019747. Now, the strain is preserved in a preservation unit (the name of the preservation unit is China center for type culture Collection, the preservation date is 10 months and 10 days in 2019, the preservation number is CCTCC NO: M2019747, the address of the preservation unit is Wuhan, Lojia mountain, Wuhan university and postal code 430072) appointed by the State intellectual property office.
Further, the method comprises the following steps of (1) respectively culturing four bacterial liquids: preparing beef extract peptone liquid culture medium → sterilizing → cooling → inoculating → culturing on a shaker at 200rpm for-72 hours → collecting the bacterial liquid; (2) mixing the four kinds of bacteria liquid in certain proportion; (3) application: when crops are cultivated, digging cultivation holes corresponding to the sizes of seedling roots in soil, placing seedlings, and pouring mixed bacterial liquid at the roots of the plants, wherein the using amount of the bacterial liquid is 20ml-200ml per plant according to the sizes of the roots; or, after mixing the bacterial liquid and the watering water with any proportion, watering the mixture into a cultivation hole when the crops are cultivated, or watering the mixture after covering soil.
Further, the compound nematicidal microorganism is used in the following proportion by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zoophila is mixed in a ratio of 1-2: 3-5: 1-2.
Further, the compound nematicidal microorganism is used in the following proportion by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:1:4: 1.
Further, the compound nematicidal microorganism is used in the following proportion by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:2:4: 2.
Further, the compound nematicidal microorganism is used in the following proportion by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:2:5: 1.
Further, the compound nematicidal microorganism is used in the following proportion by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 2:1:4: 1. Further, the method of the present invention comprises the step of (4) a cultivation management method of subsequent management after application: it is not possible to apply bactericidal and fungicidal agents to the roots.
Further, the method comprises the steps of a) after the bacterial liquid is cultured, adding the bacterial liquid into the sterilized wheat bran, and drying at low temperature to prepare bacterial powder; applying the dried fungal powder to a field when cultivating plants; or activating the fungus powder in the nutrient solution, and pouring the activated fungus powder into the roots of the plants during cultivation;
or b) freeze-drying the mixed bacterial liquid to prepare pure bacterial powder, and directly applying the dried bacterial powder when the plant is cultivated, or applying the bacterial powder after mixing with other substances, or pouring the bacterial liquid containing activated bacteria on the root of the plant after the bacterial powder is activated.
The invention has the beneficial effects that 1) the Burkholderia arabica is a strain obtained by new research, and the strain has higher prevention and treatment effect on nematodes, and new functions are in continuous research; 2) on the basis of preventing and treating the nematodes by the burkholderia arabica, the other three strains are mixed, and the related volume ratio is controlled, so that the results of outstanding preventing and treating effects and high preventing and treating rate are realized [ see an embodiment ]; 3) the compound nematicidal microorganism has the capability of obviously improving the nematodiasis resistance of field crops; meanwhile, the commodity quality and the yield of the crops can be improved.
The invention is further explained below with reference to the drawings and the detailed description.
Drawings
FIG. 1 is a diagram of the prevention of root damage of tobacco inoculated with nematodes by single strains and mixed bacteria according to the potting experiment of the present invention.
FIG. 2 is a statistical chart of agronomic characteristics-plant height of tobacco after the tobacco field uses each single microbial inoculum and each compound microbial inoculum.
FIG. 3 is a statistical chart of agronomic characteristics-stem circumference of tobacco after the tobacco field uses each single microbial inoculum and each composite microbial inoculum.
FIG. 4 is a statistical chart of the agronomic characteristics of tobacco, namely the maximum leaf length, after the tobacco field uses each single microbial inoculum and each composite microbial inoculum.
FIG. 5 is a statistical chart of the agronomic characteristics of tobacco, namely the maximum leaf width, after the tobacco field uses each single microbial inoculum and each composite microbial inoculum.
FIG. 6 is a statistical chart of the agronomic characteristics-maximum leaf area of tobacco using single and complex bacterial agents in tobacco fields.
Detailed Description
A method for preventing and treating plant nematode diseases by using composite nematocidal microbe features that four different microbial strains with nematocidal function, including gliocladium roseum (or gliocladium roseum)Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyces lilacinus(Thom.) Samson), Burkholderia arabica (B.arabic)Burkholderia arboris) Serratia zoophila (A), (B), (CSarratia entomophilus) The composite microbial inoculum prepared by the method has obvious effect of preventing and treating plant nematode diseases through field experiments.
The method for preventing and treating plant nematode diseases by using the compound microbial inoculum comprises the following steps of:
firstly, respectively culturing 4 bacterial liquids: preparing beef extract peptone liquid culture medium → sterilizing → cooling → inoculating → culturing for 12-72 hours by 200rpm of a shaking table → collecting the bacterial liquid.
Secondly, mixing the 4 bacterial liquids according to the proportion of gliocladium roseum: (Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyces lilacinus(Thom.) Samson), Burkholderia arabica (B.arabic)Burkholderia arboris) Serratia zoophila (A), (B), (CSarratia entomophilus) The volume ratio of the mixture is 1:1:4:1 (v: v: v: v).
Third, bacterial liquid application
When the crops are cultivated, the cultivation holes corresponding to the sizes of the roots of the seedlings are dug in the soil, the seedlings are placed, the mixed bacterial liquid is poured on the roots of the plants, the using amount of the bacterial liquid is 20ml-200ml per plant according to the sizes of the roots, or the bacterial liquid and water for pouring the roots in any proportion can be mixed and then poured into the cultivation holes when the crops are cultivated, or the mixture is covered with soil and then poured.
Fourth, management after administration
And (3) a subsequent cultivation management method: the management is carried out according to the conventional cultivation management mode of applying crops, but medicines such as bactericides, fungicides and the like cannot be applied to roots.
The invention can add the bacteria liquid into the sterilized wheat bran after the bacteria liquid is fermented, and the bacteria powder is prepared after the bacteria liquid is dried at low temperature. In the cultivation of plants, the dried fungal powder is applied to the field. The fungal powder can also be activated in nutrient solution and poured into the root of plant during cultivation.
The mixed bacteria liquid can be freeze-dried to prepare pure bacteria powder, and the dried bacteria powder is directly applied when the plant is cultivated, or the bacteria powder is mixed with other substances and then applied, or the bacteria powder is activated and then the bacteria liquid containing activated bacteria is poured on the root of the plant.
Besides the application of the microbial inoculum of the invention during the cultivation, the fermented bacteria liquid can be irrigated at the root part in the growth period after the plant cultivation to achieve the purpose of killing the nematodes.
The product can also be used in combination with other nematode biocontrol microbial inoculum to further enhance the control effect of the microbial inoculum.
Detailed description of the preferred embodiment
And (5) finishing the tobacco field soil according to the conventional requirement. Selecting strong and tidy tobacco seedlings with the height of about 10cm, the plant distance of 50cm and the pit depth of 15 cm. In each hole for cultivating the tobacco seedling, 20ml of mixed bacterial liquid is applied firstly, and then the tobacco is managed by measures such as earthing, watering and the like according to the tobacco seedling cultivation standard, so that the tobacco seedling can grow normally. The effect of nematode control can be seen after the tobacco plants grow.
Detailed description of the invention
Selecting tomato seedlings growing normally, digging small holes beside roots and making the holes deep to lateral roots. And applying 50ml of mixed bacteria liquid into each hole, then covering soil, and then managing according to a normal vegetable seedling management method, so that the vegetable seedlings can grow normally.
Detailed description of the preferred embodiment
And (5) finishing the tobacco field soil according to the conventional requirement. Selecting strong and tidy tobacco seedlings with the height of about 10cm, the plant distance of 50cm and the pit depth of 15 cm. 100ml of mixed bacterial liquid is applied to each hole for cultivating the tobacco seedlings, and then the tobacco is managed by measures such as soil covering, watering and the like according to the tobacco seedling cultivation standard, so that the tobacco seedlings can grow normally. The effect of nematode control can be seen after the tobacco plants grow.
Detailed description of the invention
And (5) finishing the tobacco field soil according to the conventional requirement. Selecting strong and tidy tobacco seedlings with the height of about 10cm, the plant distance of 50cm and the pit depth of 15 cm. In each hole for cultivating the tobacco seedling, 1 g of the freeze-dried bacterium powder of the mixed bacterium liquid is dissolved in 1L of root watering water, and then the tobacco is managed by measures such as soil covering, watering and the like according to the tobacco seedling cultivation standard, so that the tobacco seedling grows normally. The nematode control effect can be seen after the tobacco plant grows.
Detailed description of the preferred embodiment
And (5) finishing the tobacco field soil according to the conventional requirement. Selecting strong and tidy tobacco seedlings with the height of about 10cm, the plant distance of 50cm and the pit depth of 15 cm. Then, the tobacco is managed by measures such as soil covering, watering and the like according to the tobacco seedling cultivation standard, and the tobacco seedlings can grow normally. In each hole for cultivating tobacco seedlings, 20ml of mixed bacterial liquid is applied firstly, after the tobacco plants are fixedly planted, 10ml of bacterial liquid is diluted by 90ml of clear water each time and then irrigated to roots in three different periods, and the rest is managed according to the normal management method of the tobacco plants.
In FIG. 1, Ck0 is a group of missed nematodes, CK1 is a group inoculated with nematodes but without any nematicide, SX1 is a group of Gliocladium roseum, SX2 is a group of Paecilomyces lilacinus, SX3 is a group of Burkholderia cepacia, strain SX4 is a group of Serratia zoophila, and composite is a group of Gliocladium roseum (A), (B), (Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyces lilacinus(Thom.)Samson) Burkholderia arabidopsis: (A. arabinosus)Burkholderia arboris) Serratia zoophila (A), (B), (CSarratia entomophilus) Is a mixed microbial inoculum formed by mixing the components in a volume ratio of 1:1:4:1 (v: v: v: v).
In fig. 2 to 6, CK 0: blank control; CK 1: conventional control, conventional medicament control root knot nematode disease; t1: 5mL of Burkholderia arabica per tobacco plant; t2: 25mL of Burkholderia arabica per tobacco; t3: 50mL of Burkholderia arabinonii per strain of tobacco; t4: 25mL of serratia nematophila per strain of tobacco; t5: 20g of paecilomyces lilacinus bacterial manure per plant; t6: 20g of gliocladium roseum bacterial manure per plant; t7: each strain of paecilomyces fumosoroseus bacterial manure, gliocladium roseum bacterial manure, burkholderia arabii and serratia nematophila; t8: luofuda agent; t9: a fosthiazate chemical agent. The T7 complex microbial inoculum shows the best effect.
The above description is only a part of specific embodiments of the present invention (since the formula of the present invention belongs to the numerical range, the embodiments are not exhaustive, and the protection scope of the present invention is subject to the numerical range and other technical point ranges), and the detailed contents or common knowledge known in the schemes are not described too much. It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by means of equivalent substitution or equivalent transformation for those skilled in the art are within the protection scope of the present invention. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (9)
1. A method for preventing and eliminating nematode disease by using composite nematocidal microbe is characterized by that said composite nematocidal microbe is prepared from gliocladium roseum (Gliocladium roseum) (Zusamii)Gliocladium roseum) Paecilomyces lilacinus (A) and (B)Paecilomyces lilacinus(Thom.) Samson), Burkholderia arabica (B.arabic)Burkholderia arboris) Serratia zoophila (A), (B), (CSerratia entomophilus) Four components; the Burkholderia arabidopsis isBurkholderia arborisJ211; the preservation number is CCTCC NO: m2019747.
2. The method for controlling nematode diseases using composite nematicidal microorganisms according to claim 1, wherein the method comprises the steps of (1) separately culturing four bacterial solutions: preparing beef extract peptone liquid culture medium → sterilizing → cooling → inoculating → culturing on a shaker at 200rpm for-72 hours → collecting the bacterial liquid; (2) mixing the four kinds of bacteria liquid in certain proportion; (3) application: when crops are cultivated, digging cultivation holes corresponding to the sizes of seedling roots in soil, placing seedlings, and pouring mixed bacterial liquid at the roots of the plants, wherein the using amount of the bacterial liquid is 20ml-200ml per plant according to the sizes of the roots; or, after mixing the bacterial liquid and the watering water with any proportion, watering the mixture into a cultivation hole when the crops are cultivated, or watering the mixture after covering soil.
3. The method for controlling nematode disease of claim 1, wherein said complex nematicidal microorganism is used in the ratio, by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zoophila is mixed in a ratio of 1-2: 3-5: 1-2.
4. The method for controlling nematode disease of claim 3, wherein said complex nematicidal microorganism is used in the ratio, by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:1:4: 1.
5. The method for controlling nematode disease of claim 3, wherein said complex nematicidal microorganism is used in the ratio, by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:2:4: 2.
6. The method for controlling nematode disease of claim 3, wherein said complex nematicidal microorganism is used in the ratio, by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 1:2:5: 1.
7. The method for controlling nematode disease of claim 3, wherein said complex nematicidal microorganism is used in the ratio, by volume: gliocladium roseum: paecilomyces lilacinus: burkholderia arabidopsis: the serratia zooophila is mixed in a ratio of 2:1:4: 1.
8. The method for controlling nematode disease with a complex nematicidal microorganism according to claim 2, further comprising the steps of (4) cultivation management methods for subsequent management after application: it is not possible to apply bactericidal and fungicidal agents to the roots.
9. The method for controlling nematode diseases using composite nematicidal microorganisms according to claim 2, wherein the method comprises the steps of a) culturing the bacterial solution, adding the bacterial solution to sterilized wheat bran, and drying the mixture at low temperature to obtain bacterial powder; applying the dried fungal powder to a field when cultivating plants; or activating the fungus powder in the nutrient solution, and pouring the activated fungus powder into the roots of the plants during cultivation;
or b) freeze-drying the mixed bacterial liquid to prepare pure bacterial powder, and directly applying the dried bacterial powder when the plant is cultivated, or applying the bacterial powder after mixing with other substances, or pouring the bacterial liquid containing activated bacteria on the root of the plant after the bacterial powder is activated.
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| CN110699305B (en) * | 2019-11-27 | 2023-01-13 | 云南大学 | Burkholderia and application thereof |
| CN113462599B (en) * | 2021-07-01 | 2022-07-15 | 云南大学 | Biological preparation containing plant source components and microorganism source components and application of biological preparation in prevention and treatment of plant nematode diseases |
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