阅读说明：本技术 应用生物媒介法防治蛀干害虫的方法 (Method for preventing and controlling stem borers by applying biological medium method ) 是由 王敦 唐汉尧 张彦龙 郑吉阳 于 2021-06-14 设计创作，主要内容包括：本发明提供了一种应用生物媒介法防治蛀干害虫的方法,该方法为：媒介生物携带昆虫病原真菌分生孢子,通过孢子粉环带后,靶向性攻击蛀干害虫,致使害虫感病死亡。本发明以个体微小,且具备高攻击能力、高搜索寄主能力的摩泽蒲螨为载体,通过林间释放装置,让摩泽蒲螨携高效昆虫病原真菌孢子自发进入蛀干害虫的蛀道,直接或间接将真菌孢子传播给蛀干害虫,使其感染真菌或被蒲螨寄生死亡。该方法不仅能够长期有效防治蛀干害虫,同时可以显著减少化学农药的使用,减少环境中的药物残留,还大幅度减少了人工劳动量。对于经济树木、林业树木和生态环境保护具有非常重要的现实意义。(The invention provides a method for preventing and controlling stem borers by applying a biological medium method, which comprises the following steps: the vector organisms carry conidia of entomopathogenic fungi, and target attack on stem borers after the conidia pass through spore powder to cause the pests to suffer diseases and die. The invention takes the pyemotes moltenuis which is tiny and has high attacking capacity and high host searching capacity as a carrier, and leads the pyemotes moltenuis to carry high-efficiency entomopathogenic fungal spores to spontaneously enter a borer of a trunk-boring pest through a forest release device, so that the fungal spores are directly or indirectly transmitted to the trunk-boring pest, and the trunk-boring pest is infected with fungi or parasitized by the pyemotes. The method can effectively prevent and treat the trunk borers for a long time, can obviously reduce the use of chemical pesticides, reduce the drug residues in the environment and greatly reduce the amount of manual labor. Has very important practical significance for economic trees, forestry trees and ecological environment protection.)

1. A method for preventing and controlling stem borers by applying a biological medium method is characterized by comprising the following steps: the vector organisms carry conidia of entomopathogenic fungi, and target attack on stem borers after the conidia pass through spore powder to cause the pests to suffer diseases and die.

2. The method of claim 1 for controlling stem-boring pests using a biological agent process, wherein: the vector organisms are the cattail mites processed by the nano materials.

3. The method of claim 1 for controlling stem-boring pests using a biological agent process, wherein: the conidium of the entomopathogenic fungi is obtained by culturing the entomopathogenic fungi on a Sachs culture medium until the conidium is mature, and collecting the conidium.

4. A method of controlling stem-boring pests using a biological agent according to claim 3, wherein: the entomopathogenic fungi are isolated from trunk-boring pests which naturally infect the fungi.

5. The method for controlling stem-boring pests using a biological agent according to any one of claims 1 to 4, wherein: the method is realized through release device, release device includes centrifuging tube (1), the open-top setting of centrifuging tube (1), a plurality of have been placed to the bottom of centrifuging tube (1) and have produced ripe suge pyemotes and expand the abdominal body individual (4), are provided with first area (2) and second area (3) on the body inner wall of centrifuging tube (1), first area (2) coating has the nano-material clitellum, and entomopathogenic fungi conidia has been placed in second area (3) to the coating has the spore powder clitellum.

6. The method of claim 5 for controlling stem-boring pests using a biological agent process, wherein: the first belt zone (2) and the second belt zone (3) are arranged at intervals, and the first belt zone (2) is positioned below the second belt zone (3).

7. The method of claim 5 for controlling stem-boring pests using a biological agent process, wherein: the coating width of the nano material ring belt is 0.1-1.5 cm; the spore powder zone has a coating width of 0.1-1.5 cm.

8. The method of claim 5 for controlling stem-boring pests using a biological agent process, wherein: the diameter of the top of the centrifugal tube is gradually reduced from bottom to top.

9. The method for controlling stem-boring pests using a biological agent according to claim 2 or 5, wherein: the nano material is nano microcrystalline chitosan.

10. The method of claim 1 for controlling stem-boring pests using a biological agent, comprising the steps of:

s1, culturing conidia of entomopathogenic fungi;

s2, preprocessing of a releasing device: placing a plurality of mature pyemotes dilatorium individuals at the bottom of the release device; a first belt area (2) and a second belt area (3) are sequentially arranged on the inner wall of the tube body of the releasing device, a nano material ring area with the width of 0.1-1.5cm is arranged on the first belt area (2), a spore powder ring area with the width of 0.1-1.5cm is arranged on the second belt area (3), and conidia of the entomopathogenic fungi are placed on the second belt area (3);

s3, fixing the releasing device processed in the step S2 on the branches of the damaged trees, and enabling the pipe openings of the releasing device to be tightly attached to the barks and to be arranged obliquely downwards according to the direction of the branches.

Technical Field

The invention belongs to the field of stem borer control, and particularly relates to a method for controlling stem borers by applying a biological medium method.

Background

The pests eating trunk are widely distributed and have various types, and are important pests of economic tree species such as forest trees, fruit trees and the like. The living places are hidden, and the prevention and treatment difficulty is high; the number of insect population is stable, and the influence of natural environment and climatic conditions is small; serious harm, influence of forest health and even death.

Such as Anoplophora glaboripennis (Motschulsky) of Anoplophora glabripennis, are distributed almost nationwide. In 1978, the 'three-north' protection forest engineering in China is destroyed, tens of thousands of trees are killed by harm, so that great economic loss is caused, and the ecological environment in the northwest is greatly destroyed.

For example, Dendroctonus armandi Tsai et Li causes large-area destructive death of Pinus warrior forest in Qinba area. By 2018, the area of the bark beetle of the Pinus armandi reaches 18511hm²。

Because of the harmful characteristics of the stem borers, no better method for completely controlling anoplophora glabripennis exists at present, so that a long-term, comprehensive and effective method for preventing and treating the stem borers is urgently needed, and the actual problem is solved.

Disclosure of Invention

In order to solve the problems and the defects of the prior art, the invention aims to provide a new technology for preventing and controlling the trunk borer by a biomedia method, and the invention is a method for specially preventing and controlling the trunk borer which has strong concealment, small influence by the external environment and difficult drug control in forestry and production.

The invention is realized by the following technical scheme:

the method for preventing and controlling the stem borers by applying the biological medium method comprises the following steps: the vector organisms carry conidia of entomopathogenic fungi, and target attack on stem borers after the conidia pass through spore powder to cause the pests to suffer diseases and die.

Further, the intermediate organism is the giant cattail mites treated by the nanometer material. The pyemotes molyticus passes through the nano material belt, and the surface area and the adhesive force of the body are increased after the body is adhered with the nano material belt, so that fungal spores are carried conveniently.

Furthermore, the conidia of the entomopathogenic fungi are obtained by culturing the entomopathogenic fungi on a Saccharomy medium until the conidia are mature, and collecting the conidia.

Further, the entomopathogenic fungi are isolated from trunk-boring pests which naturally infect the fungi (not limited to one, different strains are screened for different pest isolation).

Preferably, the entomopathogenic fungus is one of beauveria bassiana, metarhizium anisopliae or other insecticidal fungi.

The method of using biological media method prevention and cure trunk borer is realized through release, release includes the centrifuging tube, the open top of centrifuging tube sets up, and a plurality of has been placed to the bottom of centrifuging tube and has produced ripe suge pyemotes dilatopomorphia individual, is provided with first area and second area on the body inner wall of centrifuging tube, first area coating has the nano-material clitellum, and entomopathogenic fungi conidium has been placed to the second area to the coating has the spore powder clitellum.

Further, the first belt and the second belt are arranged at intervals, and the first belt is positioned below the second belt.

As a further preferable scheme, the coating width of the nano-material annular belt is 0.1-1.5 cm; the spore powder zone has a coating width of 0.1-1.5 cm.

As a further preferred scheme, the diameter of the top of the centrifugal tube is gradually reduced from bottom to top.

As a further preferable scheme, the nano material is nano microcrystalline chitosan.

The method for preventing and controlling the stem borers by applying the biological medium method comprises the following steps:

s1, culturing conidia of entomopathogenic fungi;

s2, preprocessing of a releasing device: placing a plurality of mature pyemotes dilatorium individuals at the bottom of the release device; sequentially arranging a first belt area and a second belt area on the inner wall of a pipe body of the release device, arranging a nano material ring area with the width of 0.1-1.5cm in the first belt area, arranging a spore powder ring area with the width of 0.1-1.5cm in the second belt area, and placing conidia of the entomopathogenic fungi in the second belt area;

s3, fixing the releasing device processed in the step S2 on the branches of the damaged trees, and enabling the pipe openings of the releasing device to be tightly attached to the barks and to be arranged obliquely downwards according to the direction of the branches.

As a further preferable scheme, the acquisition path of the conidium of the entomopathogenic fungi cultured by the S1 can adopt commercial insecticidal fungal spores, and can also be obtained by self-separation in the following way:

s101, bringing the trunk borers infected with fungi from a natural state back to a laboratory, placing the trunk borers on an aseptic operation table, disinfecting the surface of the trunk borers by using 75% alcohol, cutting the trunk borers, taking a small amount of infected tissues inside, inoculating the trunk borers on a PDA culture medium, and placing the trunk borers in a biochemical incubator at 25 ℃ for 10 days; picking up the grown hyphae, and inoculating the hyphae to a new PDA culture medium again, and performing purification culture under the above conditions; after three rounds of purification and culture are repeated, the entomopathogenic fungi are obtained by separation; observing the shapes of hypha and conidium of the obtained entomopathogenic fungi through a microscope, performing molecular identification by adopting corresponding conservative gene primers, and storing the strains after identifying without errors.

S102, carrying out sporulation on the entomopathogenic fungi obtained in the step S101 on 1/4 Sachs medium at the temperature of 20-25 ℃ and the relative humidity of 40-60%, and scraping conidia to a centrifugal tube for later use by using a brush after 15-30 days.

Due to the adoption of the technical scheme, the invention has the following beneficial effects:

(1) long-term green and efficient prevention and treatment: the invention takes the pyemotes molyticus as a carrier, and leads the pyemotes molyticus to carry high-efficiency entomopathogenic fungal spores to spontaneously enter a borer of a trunk-boring pest through a simple forest release device, and directly or indirectly spread the fungal spores to the trunk-boring pest, so that the trunk-boring pest is infected with fungi or parasitized by the pyemotes.

(2) The application is convenient: the mature pyemotes moxystus majus puffer and the simple releasing device of the conidia of the entomopathogenic fungi are fixed in the forest.

(3) The invention can achieve the control effect through a plurality of factors, and the trunk borers are mainly infected with the fungus and killed by contacting the entomopathogenic fungal spores transmitted into the wormhole by the pyemotes molesta; meanwhile, the possibility that the trunk borers are directly parasitized and killed by the Tyrophagus molesta is also existed; or the pyemotes molezans and the entomopathogenic fungi kill the stem-borers together; particularly, the entomopathogenic fungi grow and colonize the wormhole sufficiently to form continuous infection on stem borers.

(4) The medium organisms adopted by the invention are mites of parasitic insects, and the medium organisms are small and easy to enter the insect channels of trunk-boring pests.

(5) The host of the entomopathogenic fungi adopted by the invention is insects, is safe to higher animals, does not pollute the environment and is harmless to people and livestock.

(6) The release method adopted by the invention is simple to operate and low in cost, and compared with other control methods, the manual labor is greatly reduced; the invention also has the characteristics of comprehensive, long-term, convenient and green prevention and control of the stem borers, and is easy to popularize and apply in a large scale.

The foregoing description is only an overview of the technical solutions of the present invention, and in order to clearly understand the technical solutions of the present invention and to implement the technical solutions according to the contents of the description, the following detailed description is given with reference to the preferred embodiments of the present invention and the accompanying drawings.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other designs and drawings can be obtained based on these drawings without creative efforts.

Fig. 1 is a schematic structural view of the release device of the present invention.

Detailed Description

In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.

It should be noted that the implementation conditions used in the examples can be further adjusted according to the specific experimental environment, and the implementation conditions not mentioned are generally the conditions in the routine experiments. The preparation methods mentioned in the invention are all conventional methods unless otherwise specified.

All starting materials mentioned in the following examples are obtained from published commercial sources unless otherwise specified.

The invention provides a method for preventing and treating stem borers by applying a biological medium method, which comprises the following steps: the vector organisms carry conidia of entomopathogenic fungi, and target attack on stem borers after the conidia pass through spore powder to cause the pests to suffer diseases and die. Specifically, the vector organisms adhere to the nano material and the conidium powder of the entomopathogenic fungi in the release process, actively search for a moth hole to transmit the fungal spore into a moth channel of trunk-boring pests, and effectively infect the trunk-boring pests in a targeted manner.

Further, the intermediate organism is the giant cattail mites treated by the nanometer material.

Furthermore, the conidia of the entomopathogenic fungi are obtained by culturing the entomopathogenic fungi on a Saccharomy medium until the conidia are mature, and collecting the conidia.

The entomopathogenic fungi are separated from stem borers which are infected with the fungi in a natural state, have high infectivity on the stem borers and do not pollute the environment. The strain is not limited to a species, and is selected depending on the species of the trunk-boring pest. The entomopathogenic fungi are adhered to the mite body through the spore powder belt to form an infection source, and the mite carries conidium to search pests.

The method of using biological media method to prevent and treat trunk borer is realized through release, as shown in fig. 1, release includes centrifuging tube 1, centrifuging tube 1's open top sets up, and a plurality of has been placed at the bottom of centrifuging tube 1 and has produced ripe pyemotes dilatopomorphia individual 4, is provided with first band region and second band region 3 on the body inner wall of centrifuging tube 1, proper amount vaseline is paintd in first band region, then will paint the nano-material that the width is 0.1-1.5cm on vaseline, form the nano-material clitellum, insect pathogenic fungus conidia has been placed to second band region 3 to paint proper amount vaseline on second band region 3, then will paint the spore powder that the width is 0.1-1.5cm on vaseline, form spore clitellum powder. The son mites generated by the pyemotes moxibunda puffiness adhere to the nano material through the nano material girdle, so that the spore adhesion capability is improved; after the sub-mites pass through the spore powder ring, conidia are carried on mite bodies, the sub-mites are used as vector organisms to transmit entomopathogenic fungi while seeking hosts, and the pest wormholes are attacked in a targeted mode, so that the pest is infected and killed.

Further, the first belt zone 2 and the second belt zone 3 are arranged at intervals, and the first belt zone 2 is located below the second belt zone 3.

As a further preferred scheme, the diameter of the top of the centrifugal tube 1 is gradually reduced from bottom to top.

As a further preferable scheme, the nano material is nano microcrystalline chitosan.

The method for preventing and controlling the stem borers by applying the biological medium method comprises the following steps:

s1, culturing conidia of entomopathogenic fungi;

s2, preprocessing of a releasing device: placing a plurality of mature pyemotes dilatorium individuals at the bottom of the release device; sequentially arranging a first belt area 2 and a second belt area on the inner wall of a pipe body of the release device, arranging a nano material ring area with the width of 0.1-1.5cm in the first belt area 2, arranging a spore powder ring area with the width of 0.1-1.5cm in the second belt area, and placing conidia of the entomopathogenic fungi in the second belt area;

s3, fixing the releasing device processed in the step S2 on the branches of the damaged trees, and enabling the pipe openings of the releasing device to be tightly attached to the barks and to be arranged obliquely downwards according to the direction of the branches.

As a further preferred embodiment, the S1 culturing conidia of entomopathogenic fungi comprises:

s101, bringing the trunk borers infected with fungi from a natural state back to a laboratory, placing the trunk borers on an aseptic operation table, disinfecting the surface of the trunk borers by using 75% alcohol, cutting the trunk borers, taking a small amount of infected tissues inside, inoculating the trunk borers on a PDA culture medium, and placing the trunk borers in a biochemical incubator at 25 ℃ for 10 days; picking up the grown hyphae, and inoculating the hyphae to a new PDA culture medium again, and performing purification culture under the above conditions; after three rounds of purification and culture are repeated, the entomopathogenic fungi are obtained by separation; observing the shapes of hypha and conidium of the obtained entomopathogenic fungi through a microscope, performing molecular identification by adopting corresponding conservative gene primers, and storing the strains after identifying without errors;

s102, carrying out sporulation on the entomopathogenic fungi obtained in the step S101 on 1/4 Sachs medium at the temperature of 20-25 ℃ and the relative humidity of 40-60%, and scraping conidia to a centrifugal tube 1 for later use by a brush after 15-30 days.

The invention takes the pyemotes molticus with excellent characteristics of high attack ability, high search ability and the like as a carrier, and leads the pyemotes molticus to carry high-efficiency entomopathogenic fungal spores to spontaneously enter the wormhole of the stem-borer through a simple release device, directly or indirectly spread the fungal spores to the stem-borer, so that the stem-borer is infected with the fungi or parasitized and died by the pyemotes, thereby achieving the effect of comprehensively, conveniently and greenly preventing and treating the stem-borer for a long time.

It is to be noted that the mediator biological pyemotes mimicus is provided by a forest ecological environment and protection research institute of China forestry science research institute, Zhang Yanlongong.

The nano material is nano microcrystalline chitosan.

The entomopathogenic fungi described in the following examples are Beauveria bassiana obtained by culturing larvae of longicorn infected with the fungi in a natural state on a separation medium. Sporulation is carried out on 1/4 Sachs culture medium at the temperature of 20-25 ℃ and the relative humidity of 40-60%, and conidia are scraped into a 1.5mL centrifugal tube for standby after 15-30 days.

Example 1:

as shown in figure 1, 1 mature pyemotes moxystus individual is placed at the bottom of the simple release device; the nanometer material is placed in a first zone 2, and the conidia of the beauveria bassiana is placed in a second zone, so that a nanometer material belt with the width of 0.2cm and a spore powder ring belt are respectively formed.

Example 2:

5 mature pyemotes moxystus individual bodies are placed at the bottom of the simple release device; the nanometer material is placed in a first zone 2, and the conidia of the beauveria bassiana is placed in a second zone, so that a nanometer material belt with the width of 0.5cm and a spore powder ring belt are respectively formed.

Example 3:

50 mature pyemotes moxystus individual bodies are placed at the bottom of the simple release device; the nanometer material is placed in a first zone 2, and the conidia of the beauveria bassiana is placed in a second zone, so that a nanometer material belt with the width of 1.0cm and a spore powder girdle are respectively formed.

Determination of controlling Effect

In the embodiments 1-3 of the novel technology for preventing and controlling stem borers by the biological medium method, anoplophora glabripennis and dendroctonus valens beetles are respectively used as the objects to be prevented and controlled. Fixing a releasing device carrying mature pyemotes moxibunda dilatant and conidia of entomopathogenic fungi on branches and stems of elms seriously harmed by anoplophora glabripennis by using a plastic package strip, slightly inclining the pipe orifice downwards, and splitting the elms after 15-30 days to check the control effect; and winding and fixing the releasing device carrying the mature pyemotes moxibunda turgida and conidia of the entomopathogenic fungi on the trunk of the pinus armandii with serious damage of the bark beetles of the pinus armandii by using gauze, slightly inclining the pipe orifice downwards, and stripping off the bark or splitting the tree body after 15-30 days to check the control effect.

(1) The invention relates to a new technology for preventing and controlling stem borers by a biological medium method, which comprises the following steps:

the prevention and treatment technology comprises the following steps: the pyemotes mollis treated by the nano material is taken as a carrier, and the pyemotes mollis carrying entomopathogenic fungi spores spontaneously enters a moth channel of a trunk-boring pest through a forest release device.

Trees damaged by trunk borers, 3 trees were treated per group as follows:

a) control group 3:

the prevention and cure are not performed.

Secondly, only the releasing device is fixed on the damaged tree, and the inside of the releasing device is sterile and mite-free.

And thirdly, only the releasing device is fixed on the damaged tree, and the inside of the releasing device is sterile and only mites exist.

b) And (3) control group: the control group is 3 groups, and the new technology for controlling the stem borers by the biomedia method of the embodiment 1 to 3 is respectively adopted in sequence. The victim tree is fixed with a releasing device which is loaded with mature pyemotes moxystus mazeri puffiness and conidia of entomopathogenic fungi.

The distribution of the releasing devices is determined according to the number of trees in a test field, and one releasing device is fixed at intervals of 1 tree; the treatment group comprises forest lands of six different test points, and each forest land is fixed with 5 release devices to control 10 trees.

c) After 15-30 days of new technology for preventing and controlling stem borers by a biological medium method, respectively dissecting the stems of the control trees one by one to check the death number and the live insect number of anoplophora glabripennis; stripping off the bark to check the death number and the number of live insects of the ips manillensis, counting the death rate of the anoplophora glabripennis or the ips manillensis, and calculating the average value.

(2) The biological medium method of the invention prevents and treats the new technological prevention and cure effect of the trunk borer:

aiming at the serious damage of anoplophora glabripennis and the dendroctonus valens in forestry, after one month of new technology for preventing and controlling the stem-borers by a biomedia method, the death rates of the two stem-borers are respectively calculated by forest time investigation, and the results are shown in tables 1 and 2.

TABLE 1 control of mortality in anoplophora glabripennis

Treatment method	Average mortality/%)
		Example 1 New technique for controlling trunk-boring insects by biological medium method	90.1
Example 2 New technique for controlling trunk-boring insects by biological medium method	89.2
		Example 3 New technique for controlling trunk-boring insects by biological medium method	92.7
Comparison: sterile and acarid-containing	15.6
		Comparison: sterile acarus-free	0
Comparison: do not prevent and cure	0

Mortality of Dendrolimus punctatus

Treatment method	Average mortality/%)
		Example 1 New technique for controlling trunk-boring insects by biological medium method	89.2
Example 2 New technique for controlling trunk-boring insects by biological medium method	95.4
		Example 3 New technique for controlling trunk-boring insects by biological medium method	92.1
Comparison: sterile and acarid-containing	17.3
		Comparison: sterile acarus-free	6.3
Comparison: do not prevent and cure	5.5

The results show that: the examples 1-3 all produce good control effect on the stem borers, compared with the contrast, the new technology for preventing the stem borers by the biological medium method produces very strong control effect, and the mortality rate of the anoplophora glabripennis and the larch beetles after control is more than 90 percent on average.

The prevention and treatment effect shows that the novel technology for preventing and treating the stem borers by the biological medium method has obvious effect on preventing and treating the stem borers in forestry. Because the traditional chemical control method is difficult to contact the stem borers hidden in living places and has large manual labor amount for controlling. The novel technology for preventing and treating the stem borers by the biological medium method not only can effectively prevent and treat the stem borers for a long time, but also can obviously reduce the use of chemical pesticides and reduce the drug residues in the environment, and in addition, the amount of manual labor is greatly reduced. Has very important practical significance for economic trees, forestry trees and ecological environment protection.

In conclusion, the invention utilizes the pyemotes molesta which is tiny and has high attacking capacity and high host searching capacity, simultaneously applies the entomopathogenic fungi separated from the trunk-boring pests infected with the fungi in the natural state, and combines a plurality of biological control approaches to effectively control the trunk-boring pests in a green way.