CN115669434A - Method for cooperatively treating Chinese chive maggots by low oxygen stress and entomopathogenic nematodes - Google Patents
Method for cooperatively treating Chinese chive maggots by low oxygen stress and entomopathogenic nematodes Download PDFInfo
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Abstract
The invention discloses a method for processing Chinese chive maggots by cooperation of hypoxia stress and entomopathogenic nematodes, which comprises the following steps: step 1, submerging Chinese chive maggots by using a low-oxygen reagent for pretreatment, wherein the concentration of dissolved oxygen in the low-oxygen reagent is less than 4mg/L; and 2, continuously processing the Chinese chive maggots in a manner of inoculating entomopathogenic nematodes. The water flooding pretreatment enhances the lethality of the EPNs, the two have a synergistic effect, and the method can be applied to the treatment of the Chinese chive maggots in potted plants or field plants and has a wide application range.
Description
Technical Field
The invention relates to the technical field of insect control, in particular to a method for cooperatively treating Chinese chive maggots by hypoxia stress and entomopathogenic nematodes.
Background
The Chinese chive maggot can threaten the growth of over thirty plants and is a destructive pest for the growth of Chinese chives. At present, the method for preventing and controlling the Chinese chive maggots mainly comprises the following four methods:
first, agricultural control
Resistant varieties can be selected according to local conditions, and are subjected to crop rotation with other crops (except scallion, garlic and the like), root basking by taking off soil and soil insolation. The application of plant ash and ammonia fertilizer can loosen soil and kill Chinese chive maggot to a certain extent. We suggested that the leeks were immediately harvested and covered with a net immediately above to reduce or even prevent the strong odor of the leeks from attracting the bradysia odoriphaga adults until the wound healed and the odor disappeared and then the net was removed (for published results). The damage of the Chinese chive maggots can be reduced by irrigating the Chinese chive field with water to inhibit the activity of the Chinese chive maggots. The Chinese chives are irrigated by proper amount of decomposed biogas slurry scientifically, the egg laying environment of adults is destroyed, and the harm capability of Chinese chives maggots can be reduced.
Second, physical method
The insect-proof net can be used for isolation in the leek field, so that the invasion of adult leek maggots can be effectively prevented; yellow or black sticky boards are used for adhering imagoes, sugar and vinegar liquid and a trap lamp are used for trapping and killing Chinese chive maggot imagoes, a part of Chinese chive maggots are killed by a sun-drying high-temperature film covering method before sowing, and the method is only suitable for preventing and controlling the Chinese chive maggots when the sunshine is sufficient in summer. However, the physical means has little control effect on the Chinese chive maggots, can only realize the effect of reducing the insect pests in a short time, and is not enough to continuously control the Chinese chive maggots which occur for many generations every year.
Thirdly, chemical control
The growers often use chemical pesticides such as organic phosphorus, carbamates, neonicotinoids and the like to irrigate roots to prevent and control the Chinese chive maggots such as phoxim, imidacloprid, chlorpyrifos, beta-cypermethrin and the like. However, since the habitat and the hazardous location of the Chinese chive maggots are hidden, manufacturers may blindly increase the dosage of the agent in order to kill the Chinese chive maggots remaining in the soil to the maximum extent to maintain the yield of the Chinese chives. Moreover, pesticide is continuously and excessively used throughout the year, so that the drug resistance of the Chinese chive maggots is increased, and the drug resistance of the Chinese chive maggots to phoxim and chlorpyrifos is respectively 755 times and 696 times reported by Naja and the like.
Fourthly, biological control
At present, biological control of pests of the Eumycota family has been studied more, such as control of Lycorella auripila on mushrooms with pathogenic nematodes, control of root maggots with predatory mites, etc. The biological prevention and treatment measures of the Chinese chive maggots comprise the use of microorganisms such as matrine, abamectin, bacillus thuringiensis, beauveria bassiana, bacillus subtilis and the like. In recent years, researchers have tried to utilize entomopathogenic nematodes to biologically control Chinese chive maggots, and the Chinese chive maggots are released before the Chinese chive maggots are damaged (spring equinox and autumn equinox in the north) to cause the Chinese chive maggots to die from septicemia. However, when plant-source or microorganism-source pesticides are used for preventing and controlling Chinese chive maggots, the high production cost and the relatively complex soil biological and non-biological environments become main limiting factors for popularization and application.
Disclosure of Invention
The invention aims to provide a method for cooperatively treating Chinese chive maggots by low oxygen stress and entomopathogenic nematodes aiming at the technical problems in the prevention and control of the Chinese chive maggots.
The technical scheme adopted for realizing the purpose of the invention is as follows:
the method for processing the Chinese chive maggots by the cooperation of hypoxia stress and entomopathogenic nematodes comprises the following steps:
and 2, continuously processing the Chinese chive maggots in a manner of inoculating entomopathogenic nematodes.
In the above technical solution, the pretreatment time in step 1 is 1.5 to 2.5 hours, preferably 2 hours.
In the above technical solution, the entomopathogenic nematode in step 2 is a nematode of strongyloides besseyi or heterorhabditis spp.
In the above technical solution, the concentration of the dissolved oxygen is lower than 2mg/L, and further, the concentration of the dissolved oxygen is lower than 1.5mg/L.
In the above technical solution, the hypoxia reagent includes water and an oxygen scavenger.
In the above technical solution, the oxygen scavenger is one or more of pyrosulfite and sulfite, the pyrosulfite is preferably sodium pyrosulfite or potassium pyrosulfite, the sulfite is preferably sodium sulfite or potassium sulfite, and more preferably, the oxygen scavenger is sodium pyrosulfite.
In the above technical solution, the mass percentage of the oxygen scavenger in the hypoxia reagent is greater than 0 and less than 10%, preferably 0.1-1%, and more preferably 0.4-1%.
In the technical scheme, when the Chinese chive maggots in the soil pot are treated, the soil pot planted with plants is soaked by the hypoxia reagent, and then the entomopathogenic nematodes are inoculated into the soil pot.
In the technical scheme, when the chive maggots in the field are treated, soil with the depth of 5-10cm on the ground surface is collected, the soil is soaked by a hypoxia reagent, the soil is backfilled to the field, and entomopathogenic nematodes are inoculated in the field; collecting soil 5-10cm above ground surface, soaking in hypoxia reagent, inoculating entomopathogenic nematode, treating for 12-60 hr, and backfilling into field.
In another aspect of the invention, the use of a combination of hypoxia stress and entomopathogenic nematodes for controlling insects.
Compared with the prior art, the invention has the beneficial effects that:
1. the research of the application finds that the dissolved oxygen concentration and the water flooding duration can influence the survival of the chive maggots and the entomopathogenic nematodes, and the death number of the chive maggots and the entomopathogenic nematodes is obviously increased along with the reduction of the dissolved oxygen concentration.
2. The plate test results of carrying out water-logging pretreatment on the tested Chinese chive maggots and then inoculating entomopathogenic nematodes show that the death number of the Chinese chive maggots is increased by the water-logging pretreatment and the addition of the entomopathogenic nematodes, and the dissolved oxygen concentration and the species of the entomopathogenic nematodes have obvious influence on the death number of the Chinese chive maggots subjected to the water-logging pretreatment.
3. The water flooding pretreatment enhances the lethal ability of entomopathogenic nematodes, the two have a synergistic effect, and the method can be applied to treatment of Chinese chive maggots in potted plants or field plants and has a wide application range.
Drawings
FIG. 1 is a graph of the effect of dissolved oxygen concentration on mortality of Hb nematodes;
FIG. 2 is a graph of the effect of dissolved oxygen concentration on mortality of Sf nematodes;
FIG. 3 shows the control effect of chive maggots by combining hypoxia flooding pretreatment with entomopathogenic nematodes; EPN-means not inoculated EPN, sf means inoculated Steinernema feliae, hb means inoculated Heterorhabditis bacteriophora. A-J represent 10 survey results from 12h to 120 h. CK represents no flooding.
FIG. 4 is the effect of different sulfite hypoxic stress on Bradysia odoriphaga;
figure 5 is the effect of different concentrations of sodium metabisulfite on Chinese chive maggots.
The dispersed oxygen in the figure is the Dissolved oxygen concentration, and the flooding time is the flooding time;
EPN (entomopathogenic nematodes) in the attached figures are entomopathogenic nematodes.
Detailed Description
The present invention will be described in further detail with reference to specific examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not delimit the invention.
Experimental Material
Bradysia odoriphaga Yang et Zhang: the root system of the Chinese chives damaged by the Chinese chives maggots is dug from a green Chinese chives experimental base in Tianjin Wuqing district and brought back to the laboratory. Preparing a plurality of 9cm disposable culture dishes, putting a layer of circular filter paper sheets in the culture dishes, dripping 10ml of sterile water to maintain humidity, adding 3 sections of Chinese chives in the culture dishes, picking out the Chinese chive maggots from the roots or the bulbs of the damaged Chinese chives with a small brush, and putting the Chinese chives maggots in the prepared culture dishes for later use.
Entomopathogenic nematodes EPNs: the nematode Steinernema feliae (Sf) and Heterorhabditis bacteriophora (Hb) were gifted by doctor David, ministry of agriculture and research (USDA-ARS) of the United states department of agriculture, and were bred in the laboratory. Preparing a 24-pore plate, placing a healthy and viable larva of galleria mellonella in each pore, taking the EPNs solution to estimate the density of the larva, ensuring that about 100IJs is added into each pore, and placing the pores in a constant-temperature dark incubator for culture at 25 ℃. And (3) after about 48 hours, the greater wax moth dies, the greater wax moth corpses are transferred to a White trap device after two days, the nematode solution in a large culture dish is collected for about one week, the nematode solution is washed for 3 times by using sterilized water and then transferred to a cell culture bottle, and the nematode solution is stored at 14 ℃ for later use.
And calibrating the dissolved oxygen electrode, wherein the calibration method refers to the specification of the PBJ-8608 type portable dissolved oxygen tester. Weighing 1g of sodium sulfite (analytically pure) and adding the sodium sulfite into 1L of distilled water to prepare 1g/L of excessive sodium sulfite mother liquor, taking 200ml of the mother liquor out of a large beaker, placing the beaker on a magnetic stirrer, placing a rotor with a proper length, opening the magnetic stirrer to adjust the rotating speed so as to enable the rotor to stably rotate in the beaker, immersing electrodes of a dissolved oxygen determinator after cleaning and zeroing sterile water into a solution to be detected (noting no bubbles exist), then slowly adding untreated distilled water into the beaker, continuously monitoring the dissolved oxygen concentration during the period until the dissolved oxygen concentration is adjusted to a target concentration and the value is stable (about 1 min), and reading data, wherein the data comprises real-time temperature, the dissolved oxygen concentration, saturation and an electrode current value.
For the preparation of a sufficient oxygen (normoxia) solution, since the dissolved oxygen concentration is temperature dependent, and the dissolved oxygen solubility in distilled water is sometimes less than 7.5mg/L, it is necessary to oxygenate distilled water with an oxygenation pump until its concentration is kept at 8.0mg/L for the experiments in the examples.
Example 1 Effect of dissolved oxygen concentration on nematodes Hb and Sf
The test includes two factors (1) dissolved oxygen concentration; and (2) soaking time.
The test was conducted in 50mL plastic cups, 80mL of the prepared solutions with dissolved oxygen concentrations of 1.5mg/L, 4.0mg/L, and 8.0mg/L were added to each cup, 10ul (10 IJs/. Mu.L) of the Hb or Sf nematode solution with good activity was added rapidly (Hb nematode solution stored in cell culture bottles was used as an absolute control), and the test apparatus was sealed with aluminum foil to prevent gas exchange between water and the outside air. The test set 6 flooding treatment times, namely 2h, 4h, 8h, 16h, 24h and 48h, and each treatment group set 5 replicates. After the treatment for the corresponding time, the death condition of Hb is counted and recorded by a microscope, and the nematodes in a stiff and paralyzed state under the microscope are dead. The whole experiment was repeated twice, for a total of 2 × 4 × 6 × 5=240 cups. The results of the effect of dissolved oxygen concentration on nematode Hb are shown in FIG. 1, and the results of the effect of dissolved oxygen concentration on nematode Hb are shown in FIG. 2.
As is clear from the results of FIG. 1, the mortality of Hb was higher in each of the treated groups than in the control group. The Hb mortality due to dissolved oxygen concentration and soaking time in both experiments showed similar trends. In test 1 and test 2, the solution dissolved oxygen concentration had a significant effect on the Hb mortality (test 1, F2,72=5.79, P =0.005 and straw 0.05; test 2, F2,72=54.8, P straw 0.001), and the immersion time also had a significant effect on the Hb mortality (test 1, F5,72=39.6, P straw 0.001; test 2, F5,72=49.1, P < 0.001). The Hb mortality increases with the decrease of the dissolved oxygen concentration, and in test 1, the Hb mortality has no difference under the conditions of the dissolved oxygen concentrations of 1.5mg/L and 4.0mg/L, but is higher than that under the condition of 8.0 mg/L; in experiment 2, the Hb mortality rate was significantly different between the dissolved oxygen concentration of 1.5mg/L and the dissolved oxygen concentration of 4.0mg/L and between the dissolved oxygen concentration of 4.5 mg/L and the dissolved oxygen concentration of 7.5 mg/L. As the flooding treatment time is extended, the mortality rate of Hb also increases gradually. In test 1, the dissolved oxygen concentration had no interaction with the immersion time (F10, 72=1.4, P = 0.197), while in the second test, the dissolved oxygen concentration exhibited interaction with the immersion time (F10, 72=4.7, P < -0.001)
As can be seen from the results in fig. 2, the Sf mortality rate was significantly higher in each treatment group than in the control group. Dissolved oxygen concentration had a significant effect on Sf mortality (F2,162 =25.3, P-slash 0.001), as well as soaking time (F5,162 =255, P < 0.001). The mortality rate of Sf increases with decreasing dissolved oxygen concentration, and the difference in the mortality rate of Sf is significant at dissolved oxygen concentrations of 1.5mg/L, 4.0mg/L and 7.5 mg/L. Along with the prolonging of the flooding treatment time, the mortality of the nematodes also gradually increases, and the mortality of the Sf after soaking for 2h, 4h, 8h, 16h, 24h or 48h is obviously different, wherein the mortality is respectively 27.7%, 40.3%, 60.1%, 75.7%, 86.7% and 93.6%. The effect of dissolved oxygen concentration and soaking time on Sf mortality also showed an interaction (F10,162 =2.4, p- <0.05).
The mortality of Sf was also significantly different between treatments (F18,189 =100.9, p-slashes 0.001) and after soaking in a solution with a dissolved oxygen concentration of 1.5mg/L for 2h, 4h, 48h, the mortality of Sf was 35.8%, 49.7%, 94.2%, respectively. When the dissolved oxygen concentration is 8mg/L, the mortality rates of Sf after soaking for 2h, 8h and 48h are respectively 23.9%, 54.4% and 89.7%.
Test results show that the solubility of dissolved oxygen and the length of time of soaking in solution both have an adverse effect on the survival of s.feliae and h.bacteriophora, and that there is an interaction between the dissolved oxygen concentration and the soaking time.
With the reduction of the dissolved oxygen concentration and the prolonging of the soaking time, the mortality rate of the two EPNs increases, but it seems that Sf is more easily influenced by the dissolved oxygen concentration and flooding, the mortality rate of Sf 35.8% is caused after the Sf is soaked in a solution with the dissolved oxygen concentration of 1.5mg/L for 2h, the mortality rate of Sf reaches 94.2% after 48h, even under the condition that the dissolved oxygen concentration is 8mg/L, the mortality rate of Sf is up to 89.7% after soaking for 48h, and the mortality rate of Hb under the same condition is only about one half of the mortality rate of Sf. The literature reports that flooding or irrigation at least needs to last for several days to achieve considerable effect of killing the Chinese chive maggots, so even if good Hb is achieved in the experiment, the low-oxygen flooding and the EPNs are not feasible to simultaneously play a role in preventing and controlling the Chinese chive maggots.
The results of example 1 illustrate that both Hb and Sf lines are intolerant to hypoxia or flooding, so we used a mode of pre-treating chive maggots with hypoxia flooding and then inoculating Hb and Sf to explore the control effect of the combination of the dissolved oxygen concentration and entomopathogenic nematodes on chive maggots.
Example 2
The assay included 2 factors: (1) four dissolved oxygen concentrations: 0. 1.5, 4 and 8mg/L; (2) entomopathogenic nematodes: without EPNs, sf, hb.
The total of 15 treatments were included:
1)0mg/L；2)0mg/L，Sf；3)0mg/L，Hb；
4)1.5mg/L；5)1.5mg/L，Sf；6)1.5mg/L，Hb；
7)4mg/L；8)4mg/L，Sf；9)4mg/L，Hb；
10)8mg/L；11)8mg/L，Sf；12)8mg/L，Hb；
there were three controls, 13) no flooding and no EPNs added; 14 ) inoculation of Sf;15 Hb inoculation).
The first step of waterflooding pretreatment was performed in a 100mL plastic cup. Randomly selecting 10 fourth-instar larvae from the reserved chive maggots picked in the culture dish, placing the larvae at the bottom of a plastic cup, then respectively adding 80ml of prepared solutions with different dissolved oxygen concentrations into each cup, and sealing the test device by using an aluminum foil to prevent the larvae from escaping and prevent the air exchange between water and the outside air. After low-oxygen stress pretreatment for 2h and 2h, removing the solution in the plastic cup, slowly transferring the Chinese chive maggots to a plastic culture dish with the diameter of 3cm and added with a piece of wet filter paper, and directly and randomly selecting 10 standby Chinese chive maggots in a control group to be placed in a plastic culture dish with the diameter of 3cm and added with a piece of wet filter paper. The amount of nematode solution inoculated was 100 pieces (20 IJs/. Mu.L). Mortality of Bradysia odoriphaga was observed at 12h, 24h, 36h, 48h, 60h, 72h, 84h, 96h, 108h and 120h after the EPNs infection. The Chinese chive maggots are touched by a small brush pen and are still dead. Each treatment and control group was set to 5 replicates and the entire experiment was repeated twice. Total 2 × 15 × 5=150 dishes. The results are shown in table 1 and fig. 3.
As can be seen from the results of table 1 and fig. 3, the mortality of the chive maggots after the pre-treatment of flooding was increased compared to the control group. The death number of the Chinese chive maggots pretreated by the solution with the dissolved oxygen concentration of 0mg/L and 1.5mg/L for 2h is not obviously different, the death rate of the Chinese chive maggots treated by 4mg/L is higher than 8mg/L, the death rate of the Chinese chive maggots treated by 36h, 48h and 84h is obviously different, and the death rate of the Chinese chive maggots treated by 4mg/L is lower than that of the Chinese chive maggots treated by 0mg/L and 1.5mg/L. Generally, in a certain range, the death number of the chive maggots increases with the decrease of the concentration of dissolved oxygen.
The mortality rate of Chinese chive maggots caused by two kinds of EPN infection is higher than that of Chinese chive maggot processed without inoculation of EPN, the addition of Sf or Hb increases the death rate of Chinese chive maggots at 10 time points for detecting the mortality rate of Chinese chive maggots in the test, sf is always obviously higher than control, and the investigation after 24 hours of Hb shows that the mortality rate of Chinese chive maggots is obviously higher than control. For comparison of Sf and Hb, four 12h-48h investigations showed that Sf has a stronger lethal effect on chive maggots than Hb, but at 60h and longer, mortality of chive maggots after Hb inoculation exceeds Sf.
After 60h, the mortality rate of the chive maggots treated by each method reaches 80% or more except that the EPNs are not added, and the death rate is not obviously improved along with the lengthening of the infection time.
Before 24h of infection, 0mg/L and 1.5mg/L solution is independently used for flooding for 2h to kill the Chinese chive maggots, the number of the Chinese chive maggots is higher than that of the Chinese chive maggots which are independently inoculated with Sf or Hb, the killing capacity of the Chinese chive maggots by single flooding treatment of 4mg/L and 8mg/L is lower than that of the single inoculated nematodes, the Chinese chive maggots can be continuously prevented and treated due to the fact that the Sf and the Hb infect the Chinese chive maggots, the death rate of the Chinese chive maggots caused by the single inoculation of the Sf exceeds that of the single hypoxia flooding treatment after 36h, the hypoxia flooding has better prevention and control in a short time, the Sf can play a role durably, and 1.5mg/L is recommended to save the use of a deoxidant. 36h investigation also finds that the death number of the chive maggots inoculated with the nematodes after the hypoxia water pretreatment of 8mg/L exceeds the effect of non-pretreatment, namely the water flooding pretreatment enhances the lethal ability of the EPNs, and the two have a synergistic effect.
TABLE 1
The concentration of dissolved oxygen and the soaking time have obvious influence on the death of the Chinese chive maggots, the two factors have interaction, and the death rate of the Chinese chive maggots after being flooded for 4 hours is 2 times of that after being flooded for 2 hours. However, in the condition of extremely low dissolved oxygen solubility even approaching to complete oxygen deficiency, the effect of the dissolved oxygen on the chive maggots is not increased along with the reduction of the oxygen concentration, for example, under the condition that the dissolved oxygen concentration is 0mg/L and 0.5mg/L, the death number of the chive maggots is not obviously different, and the same result is obtained even in the recovery experiment of the chive maggots.
The results show that the dissolved oxygen concentration and the EPN have obvious influence on the death number of Chinese chive maggots which are pre-treated in flooding, and the interaction is shown after 24 h. Firstly, the mortality rate of Chinese chive maggots subjected to flooding pretreatment is higher than that of Chinese chive maggots subjected to direct infection, and the fact that the flooding pretreatment can improve the death rate of Chinese chive maggots is ensured. Secondly, the experiment proves that the death number of the Chinese chive maggots is increased due to hypoxia stress, and in a certain range, the death number of the Chinese chive maggots is increased along with the reduction of the concentration of dissolved oxygen, for example, the death rate of the Chinese chive maggots caused by 4mg/L is always higher than 8mg/L in the experiment, and meanwhile, the death rate of the Chinese chive maggots caused by 4mg/L is obviously lower than 0mg/L and 1.5mg/L in treatment. However, we noticed that there was no significant difference between the dissolved oxygen concentrations of 0mg/L and 1.5mg/L for the number of deaths of the chive maggots at all times, i.e., after the dissolved oxygen concentration was reduced to a certain extent, the insect mortality rate no longer continued to increase with the reduction of the dissolved oxygen. Thirdly, test results prove that the death rate of the Chinese chive maggots is improved by adding the EPN, the death number of the Chinese chive maggots caused by inoculating the Sf or the Hb in the test is higher than that of the Chinese chive maggots not inoculated with the EPNs, 10 times of investigation shows that the death rate caused by the Sf is obviously higher than that of a control, the death causing effect of the Sf is better than that of the Hb when the Sf is investigated for 12-48 h, and the Hb is higher than the Sf when the infection is detected for 60h and longer, while the previous research shows that the Hb has better effect of preventing and controlling the Chinese chive maggots compared with the Sf, and the conclusion in the test is possibly related to the optimal environment for the function of the EPNs. More importantly, the combination of the two methods provides a synergistic effect compared to hypoxic flooding alone or the use of EPNs.
Studies of gene levels were performed after treatment with different dissolved oxygen concentrations, and by upregulating HIF-1 transcript levels under low and medium dissolved oxygen conditions, hypoxia stress under saturated dissolved oxygen conditions was more prone to energy compensatory responses, while hypoxic environments with low and medium dissolved oxygen resulted in upregulated expression of more antioxidant damage genes.
The agent for controlling the dissolved oxygen concentration in the flooding treatment to be 4mg/L or less can increase the expression of hypoxia inducible factor-1, and further, can increase the expression of the transcription level of hypoxia inducible factor-1 or the expression of the protein level of hypoxia inducible factor-1.
EXAMPLE 3 Effect of oxygen scavenger on Bradysia odoriphaga
Effect of four sulfite hypoxia stresses on chive maggots: respectively preparing 1% sodium metabisulfite solution, potassium metabisulfite solution, sodium sulfite solution and potassium sulfite solution. The experiment was performed in 100mL plastic cups. Randomly selecting 10 fourth-instar larvae from the standby Chinese chive maggots picked in a culture dish, placing the larvae at the bottom of a plastic cup, then respectively adding 80ml of prepared 4 sulfite solutions into each cup, adding 80ml of tap water into a control group plastic cup, sealing a test device by using an aluminum foil to prevent the larvae from escaping and prevent water from exchanging with the outside air, removing the solution after treating for 4 hours, and recovering the Chinese chive maggots in the air for 2 hours to determine the death number. Every treatment is repeated for 9 times, the chive maggots are touched by a small brush pen, and the Chinese chive maggots are still in a dead state. The whole experiment was repeated twice, for a total of 2 × 5 × 9=90 cups; the results are shown in FIG. 4. From the results shown in fig. 4, it can be seen that the effect of killing Chinese chive maggots by 1% sodium metabisulfite is the best, and the recovery rate is low in 2 hours, so that the reagent can be primarily considered as a good oxygen-removing reagent for performing low-oxygen stress on Chinese chive maggots.
This application has still designed the influence of different concentration sodium metabisulfite to the chinese chive maggot: sodium metabisulfite solutions were prepared at concentrations of 0.1%, 0.2%, 0.4% and 1%, respectively, with tap water being used as a control. The method steps are the same as above, and the results are shown in fig. 5. From the results in fig. 5, it is understood that sodium metabisulfite at a concentration of 1% is most lethal to chinese chive maggots and the recovery rate of 2 hours is the lowest.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. The method for processing the Chinese chive maggots by the cooperation of hypoxia stress and entomopathogenic nematodes is characterized by comprising the following steps of:
step 1, submerging Chinese chive maggots by using a low-oxygen reagent for pretreatment, wherein the concentration of dissolved oxygen in the low-oxygen reagent is less than 4mg/L;
and 2, continuously processing the Chinese chive maggots in a manner of inoculating entomopathogenic nematodes.
2. The method of claim 1, wherein the pretreatment time in step 1 is 1.5 to 2.5 hours, preferably 2 hours.
3. The method of claim 1, wherein the entomopathogenic nematode in step 2 is a nematode of the Steinernema or Heterodera.
4. The method of claim 1, wherein the dissolved oxygen concentration is less than 2mg/L, and further wherein the dissolved oxygen concentration is less than 1.5mg/L.
5. The method of claim 1, wherein the hypoxic agent comprises water and an oxygen scavenger.
6. The method according to claim 5, characterized in that the oxygen scavenger is one of pyrosulfite, preferably sodium metabisulfite or potassium metabisulfite, sulfite, preferably sodium sulfite or potassium sulfite, more preferably sodium metabisulfite, or a plurality of salts in any proportion.
7. The method of claim 5, wherein the oxygen scavenger is present in the hypoxic agent in an amount greater than 0 and less than 10% by weight, preferably in an amount of 0.1 to 1% by weight, and more preferably in an amount of 0.4 to 1% by weight.
8. The method of claim 1, wherein the chive maggots in the soil pot are treated by soaking the soil pot in which the plant is planted with the hypoxic agent and then inoculating the soil pot with the entomopathogenic nematodes.
9. The method of claim 1, wherein when treating chive maggots in the field, soil of 5-10cm above the ground surface is collected, soaked with the hypoxic agent, backfilled to the field, and inoculated with entomopathogenic nematodes; collecting soil 5-10cm above ground surface, soaking in hypoxia reagent, inoculating entomopathogenic nematodes, treating for 12-60 hr, and backfilling into field.
10. Use of a combination of hypoxia stress and entomopathogenic nematodes for combating insects.
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