CN108077192B - Diapause relieving technology for chrysopa perla - Google Patents
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Abstract
The invention provides a diapause relieving method for chrysopa perla, which comprises the following steps: 1) selecting chrysopa perla prepupa entering a diapause state as a diapause removing object; 2) placing the chrysopa perla prepupa selected in the step 1) in a photoperiod of 0L:24D at a temperature of 5 ℃ for more than or equal to 60 days to release diapause of the chrysopa perla; 3) transferring the premature pupation of the chrysopa perla released from diapause in the step 2) to a temperature of 26 ℃, enabling the chrysopa perla to recover growth and development under the condition of a photoperiod 16L:8D, pupating and emerging into adults, wherein the emergence rate of the adults is more than 90%, and the average emergence period is 23 days. The invention has the beneficial effects that: the diapause release time of the chrysopa perla is accurately controlled, sufficient natural enemy products are provided for biological control of various agricultural and forestry pests, the large-scale propagation of the chrysopa perla is expanded, the shelf life of the chrysopa perla products is prolonged, the control action time in the field is prolonged, the pest control efficiency is improved, and the like, so that the technical guarantee can be provided for industrial production and application of the chrysopa perla.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a diapause relieving technology for chrysopa perla.
Background
In recent years, with the change of modern agricultural cultivation modes and the comprehensive result of global climate change, the agricultural pest population structure and catastrophe law and the like all have new changes worldwide. At present, agricultural diseases and insect pests frequently outbreak in China, crop yield loss caused by the diseases and the insect pests accounts for about 10% -15% of total loss every year, and meanwhile, chemical pesticides are excessively used for disease and insect pest prevention and control for a long time, which seriously threatens food safety, ecological safety and national health in China. Therefore, the development of an environment-friendly, effective, safe and sustainable comprehensive pest control strategy has important economic, social and ecological benefits for guaranteeing national food production safety, promoting agricultural high yield and income and protecting ecological environment.
The biological control technology is an environment-friendly control strategy with sustainable development, has multiple technical advantages of simplicity, convenience, effectiveness, low energy consumption, no residue, sustainability and the like, and meets the requirement of modern agricultural development. The method for preventing and controlling the pests by using the natural enemy insects is one of the core biological prevention and control technologies, can effectively control the pests, reduce the using amount of pesticides, ensure the production safety of crops and realize sustainable development of agriculture. The technology is widely applied to developed countries such as Europe, North America and the like, and remarkable control effect and economic benefit are obtained.
At present, the main bottleneck problems limiting the large-scale production and application of natural enemy insects at home and abroad are as follows: the natural enemy insects have longer propagation period and shorter survival period, the products can not be stored for a long time, the long-distance transportation is difficult, and the like, so that the delivery time of the natural enemy products is difficult to be consistent with the occurrence period of pests, and the actual application effect is not ideal. Therefore, the method has important practical significance for regulating the development progress of the natural enemy insects. The regulation and control of the development progress of the insects can be realized, and the diapause which is the inherent genetic attribute of the insects can be utilized. By utilizing diapause of the natural enemy insects, the artificial regulation and control of the development progress of the natural enemy insects can be realized, the shelf life of products is prolonged, long-distance safe transportation is realized, the prevention and control action time of the natural enemy insects is prolonged, and the seasonal biology of the natural enemy insects can be further understood, so that the development is facilitated, and the effective pest control strategy is perfected. At present, natural enemy insect products widely applied at home and abroad basically master the technology of inducing the natural enemy insects to enter diapause, maintaining and removing the diapause, can store the produced natural enemy insects in a diapause state for a long time in time, and break the diapause before release to enable the natural enemy insects to enter a normal development state to control pests, and the diapause regulation technology becomes one of core technologies of natural enemy insect production and high-efficiency application.
Chrysopa chrysosporium fortmosa Brauer belongs to the order of the vein-wing, Chrysopa chrysosporium, mainly in Europe and Asia, and widely in provinces and cities of China. The insect is an excellent predatory natural enemy insect, can prey on various agricultural and forestry pests such as aphids, whiteflies, thrips, scale insects, liriomyza larvae, leaf mites, lepidoptera eggs, low-age larvae and the like, has the advantages of wide range of food, large food intake, high reproductive capacity, long service life, strong adaptability and the like, can prey on control objects for both larvae and adults, has lasting pest control and obvious effect, and plays an important role in the biological control of the agricultural and forestry pests. In recent years, sand flies have received a high degree of attention both at home and abroad and have been successfully used in many countries, such as the united states, the soviet union, the uk, france, canada, india, etc. In addition, the commercial and large-scale production of the chrysopa perla is realized in developed countries such as the United states, Canada, the Netherlands and the like, the chrysopa perla is widely applied in a large scale, the prevention and control effects are extremely obvious, and good economic, ecological and social benefits are obtained.
The diapause of the natural enemy insects such as the chrysopa perla is researched, the diapause insect state and the sensitive insect state of the natural enemy insects are clarified, the regulation and control effect of environmental factors (photoperiod, temperature and the like) on the diapause is clarified, the key technology for regulating, controlling, inducing, maintaining and removing the diapause of the natural enemy insects is mastered by manually regulating and controlling the environmental conditions such as photoperiod, temperature and the like, important technical support and method guidance can be provided for solving the problems of storage, transportation, application and the like of the natural enemy insect products, annual large-scale propagation of the natural enemy insects is realized, and the method has important practical significance for promoting industrial production and large-area release application of the natural enemy insects.
Disclosure of Invention
The invention overcomes the defects in the prior art, and provides a diapause relieving method for the chrysopa perla, which realizes the accurate control of the diapause relieving of the chrysopa perla, an insect which is a good predatory natural enemy, by controlling environmental factors such as temperature, illumination time, treatment time and the like and combining necessary treatment measures.
The purpose of the invention is realized by the following technical scheme.
A diapause relieving technology for chrysopa perla comprises the following steps:
1) selecting chrysopa perla prepupa entering a diapause state as a diapause removing object;
2) placing the chrysopa perla prepupa selected in the step 1) in a low-temperature environment for short-term treatment, wherein the low-temperature environment condition is that the photoperiod is (0-10) L, (14-24) D, the temperature is (0-10) DEG C, and the continuous treatment is carried out for more than or equal to 60 days;
3) transferring the chrysopa perla prepupa treated in the step 2) to a condition that the temperature is 26 ℃ and the photoperiod is 16L:8D, so that the chrysopa perla is released from diapause and eclosion into adults.
Further, the chrysopa perla diapause prepupa selected in the step 1) is a cocoon of 10 days old after cocooning.
Further, the diapause release is performed in an artificial climate box.
Further, the diapause release rate of the chrysopa perla prepupa in the step 3) is 90-95%, and the average eclosion period of the adults is 23 days.
Further, in the step 2), the photoperiod is 0L:24D, at a temperature of 5 ℃, for 60 days.
The invention has the beneficial effects that:
the diapause of the chrysopa perla prepupa is treated for 60 days at 5 ℃ in the dark, and then transferred to a normal development condition (26 ℃,16L:8D), so that more than 94 percent of individuals can be released from diapause in 23 days on average, eclosion is carried out to form imagoes, normal predation, mating, oviposition, reproduction and the like are carried out, and the pest control efficiency of natural enemy insect products is exerted. By utilizing the technology, diapause release of the chrysopa perla can be accurately controlled, sufficient natural enemy products are provided for biological control of agricultural pests, and technical guarantees are provided for large-scale propagation expanding application of the chrysopa perla, prolonging of the shelf life of the products and control action time in the field.
Detailed Description
The technical solution of the present invention is further illustrated by the following specific examples.
Experiment 1. determining diapause insect state and diapause control time of chrysopa perla
1.1 insect source for test and feeding method:
pea aphid Megoura japonica (Matsumura) is collected from Yunnan, indoor temperature is 26 ℃, photoperiod is 16L:8D (L is 16 hours in light time per day and 8 hours in dark time), broad bean seedlings are continuously fed for multiple generations as host plants, and stable population is established to be used as insect source for testing.
The Chrysopa formosa Brauer imagoes are collected from fields of corridor test bases of Chinese academy of agricultural sciences, the imagoes are placed in insect rearing cages (60 multiplied by 60cm) under the conditions of indoor temperature of 26 ℃ and light cycle of 16L:8D (normal development conditions), and pea tailbiting aphids are used as hosts to continuously raise the imagoes to establish stable populations. Under normal development conditions, new eggs are collected regularly every day (within 24 h) from the day of laying new eclosion imagoes, egg grains are placed in a feeding box (29X 20X 10cm) paved with folding filter paper strips for testing, and 60 eggs are placed in each box.
1.2 Experimental methods
In the study, all experimental conditions except for different temperature and photoperiod settings were the same in relative humidity and illumination intensity, RH70 ± 5% and illumination 8000LX, respectively.
Feeding Chrysopa perla by Pisum sativum at 26 + -1 deg.C under the condition of photoperiod 16L:8D (artificial climate box, Ningbo Jiangnan RXZ-500B), collecting Chrysopa perla eggs, and adding aphid every day after the eggs are hatched. The induction treatment experiments of high-temperature long light (L:26 ℃,16L:8D) and low-temperature short light (S:20 ℃,8L:16D) were respectively carried out in different development stages of the chrysopa perla (six stages of eggs, 1 st larva, 2 nd larva, 3 rd larva, prepupa and pupa) as shown in Table 1. After the larvae are cocooned for 5 days, the collected cocoons are transferred to 26 ℃, the cocoons are continuously observed for 20 days under the condition of 16L:8D, and the cocoons which are not eclosion are dissected under a microscope for checking the insect state and survival condition of the cocoons, and the diapause rate is counted. The number of the cocoons treated in each group is not less than 100.
TABLE 1 diapause rates of different insect attitude of chrysopa perla under high temperature long illumination (L:26 deg.C, 16L:8D) and low temperature short illumination (S:20 deg.C, 8L:16D)
Note: in the table, gray bars are diapause inducing conditions (20 ℃,8L: 16D); the white bars were under normal developmental conditions (26 ℃,16L: 8D). The larva just hatched from the egg is a larva of 1 st age, the larva of 2 st age after molting is a larva of 2 st age, the larva of 2 st age after molting is a larva of 3 st age, the pre-pupation refers to a development stage from feeding stop of an aged larva and cocooning to before molting and pupating, and the larva enters a pupation stage after molting. The six insect states of egg, 3 larval stages, prepupa and pupal stage are all whole-course induction.
1.3 determination of diapause insect status of Lissajous perna
Normally, an individual insect that has entered diapause will not continue to develop to the next insect state even if given normal development conditions for a short period of time. Therefore, according to the development period of the chrysopa perla, after the days required for completing the whole development stage from eggs to adults, after most adults are eclosion, the observation is continued for a period of time, and after the cocoons which are not eclosion are dissected, the cocoons which are not eclosion still keep the survivors, namely, the survivors are considered as diapause states, and the survival insect states are diapause insect states. Pupating all chrysopa perla larvae within 5 days after cocooning and emerging all chrysopa perla larvae into adults within 20 days under the conditions of 26 ℃,16L and 8D.
The experimental results show (Table 1) that the chrysopa perla is treated by low-temperature short illumination (20 ℃,8L:16D) at different development stages, then transferred to the normal development condition to continue to develop for 20 days, and the non-emergence individuals are all prepupa through anatomical examination. Many studies have also shown that chrysopa perla overwinter in cocoons with prepupa in northern regions of our country, which is consistent with the results of this experiment. Therefore, the diapause state of the chrysopa perla is prepupa.
1.4 judgement standard for chrysopa perla diapause
In order to ensure the accuracy and the scientificity of the chrysopa perla diapause regulation and control experiment, the experiment adopts a unified diapause judgment standard. As described above, the sand fly diapause in cocoon in a prepupa state, once it enters diapause, does not pupate even if given normal development conditions in a short period of time. All chrysopa perla larvae pupate within 5 days after cocooning under the condition of 26 ℃,16L, 8D, and the pupation mark is that black end-aged larvae molt at the lower end of cocoons can be seen through cocoon shells. According to one of the standard methods for judging diapause, namely a development history method, which is commonly used at home and abroad, if the chrysopa perla cocoons treated under different conditions are transferred to 26 ℃ and continue to develop for 10 days (2 times of the days for pupation of normal development individuals) under the condition of 16L:8D, the individuals which do not yet pupate are judged to be diapause after anatomical examination. The method is simple, convenient and easy to implement, has high accuracy, and is a reliable standard for judging chrysopa perla diapause.
1.5 determination of control time for diapause control of chrysopa perla
As shown in Table 1, the 3 rd larvae of Chrysopa perla were subjected to the critical phase of the diapause induction stimulus, and only when the 3 rd larvae were in the condition of diapause induction (S:20 ℃,8L:16D), Chrysopa perla could be induced to enter the diapause state. The diapause induction treatment is only carried out on the developmental insect states before and after the 3 th larva in different developmental stages, no diapause individual appears, the chrysopa perla cocoons can normally pupate, eclose and lay eggs, and the result is consistent with the result that all the developmental stages are under the normal development Condition (CK), and shows that the ovum stage, the 1 st larva, the 2 nd larva, the 1-2 nd larva and the prepupa are not sensitive to the induction of low-temperature short illumination. But the low-temperature short illumination has an accumulation effect on the induced stimulation effect of the larvae, except for carrying out the induced treatment on the 3 rd larvae of the chrysopa perla, if the 1 st larvae, the 2 nd larvae and/or the prepupa are simultaneously subjected to the low-temperature short illumination stimulation, the diapause generation of the prepupa is remarkably promoted, and the diapause rate can reach 100%.
According to the results of the comprehensive experiment 1, the chrysopa perla is diapaused in cocoons by prepupa, the 3 rd larva has the strongest sensitivity to the diapause induction stimulation conditions, and more diapause individuals can be obtained by diapause induction of the chrysopa perla from the low-age larva stage. Therefore, the optimal control time for diapause induction of the chrysopa perla is the whole larval stage and the prepupa stage, and 100% of chrysopa perla prepupa can be induced to enter the diapause state by regulating and controlling the photoperiod and the temperature factor.
Experiment 2 diapause induction experiment of chrysopa perla
2.1 experiment of the Effect of photoperiod and temperature on diapause Induction
In order to determine the influence of photoperiod and temperature on diapause induction, a two-factor five-level repeatability test is set, diapause induction is carried out in an artificial climate box (Ningbo south RZX-500B type), 5 photoperiod levels are respectively 8L:16D, 10L:14D, 12L:12D, 14L:10D and 16L:8D, and 3 temperature levels are respectively 18 ℃, 20 ℃ and 26 ℃.
To determine the effect of temperature on chrysopa perla diapause induction under short-light conditions, 7 temperature levels were set under short-light 8L:16D conditions as follows: 18 ℃, 20 ℃, 22 ℃, 24 ℃ and 26 ℃.
The experimental method comprises the following steps: primary eggs were collected at 26 ℃ under 16L:8D conditions (within 24 h) in the same manner as before (experiment 1, 1.1), with 3 replicates per treatment, and 60 eggs per replicate. And (3) incubating the eggs under the experimental treatment condition, adding sufficient aphids every day after the eggs are incubated, feeding until the larvae are cocooned, transferring the cocoons to a condition of 26 ℃ after the cocoons are cocooned for 5 days, continuously observing under the condition of 16L:8D, dissecting the cocoons which are not yet pupated for 10 days under a microscope to find out the survival state and the insect state, counting the total cocoon number, the pupation number, the diapause number and the death number, calculating the diapause rate, and taking the average value of 3 times of repetition as the result.
2.2 data analysis
All data in the experiment were analyzed statistically and the results are expressed as mean ± sem. Diapause rate was first transformed by arcsine square root and then analyzed for variance using ANOVA in GraphPad Prism V6.03 software, and the differential significance test was performed using Tukey's multiple comparison method.
2.3 analysis of results
(1) Effect of photoperiod on diapause Induction
Diapause rates of chrysopa perla under different photoperiodic conditions are shown in table 2. As can be seen from the table, the diapause rate of chrysopa perla prepupa is remarkably increased as the illumination time length is shortened, and the diapause type is a typical long-illumination response type.
When the temperature is 18 ℃, the diapause rate is the highest and is 100% under the condition of 8L under short illumination and 16D; when the illumination time is shorter than 14h, the diapause rate can reach more than 90 percent; however, when the photoperiod is L16: D8, the prepupa does not have diapause, and the diapause rate is 0.
The diapause induction law of the photoperiod is the same as that at 18 ℃ at 20 ℃. The diapause rate is the highest and reaches 100% under the condition of 8L to 16D short illumination; when the photoperiod is 10L:14D and 12L:12D, the diapause rate is close to 95 percent, although the diapause rate is slightly reduced, the difference with the diapause rate under 8L:16D is not obvious; when the illumination time is prolonged to 14h, the pupa prepupinating diapause rate is remarkably reduced to 63.8 percent; when the illumination is continued to be prolonged to 16h, the pupa prepuberculation rate is 0.
When the temperature is 26 ℃ and short illumination is also adopted, the diapause rate of the prepupa is higher, and when the photoperiod is shorter than 12L:12D, the diapause rate is higher than 90 percent; however, when the photoperiod is prolonged, the diapause rate is obviously reduced, and no diapause individual appears under the condition of long illumination for 16 h.
According to photoperiod response curves of the chrysopa perla at 18, 20 and 26 ℃, the critical photoperiods of the chrysopa perla at various temperatures are 14.4,14.3 and 12.9h respectively by calculating the relationship between the illumination duration and the diapause rate, and the critical photoperiods are gradually shortened along with the temperature rise.
At each temperature level, the prepupa diapause rate is higher in short illumination; and when the illumination is prolonged, the prepupa diapause rate is obviously reduced. When the photoperiod is shorter than 12L to 12D, the diapause rate at each temperature is maintained at a higher level, which reaches more than 90 percent; when the photoperiod is prolonged to 14L:10D, the diapause rate is obviously reduced; the diapause is not induced at all under the long illumination of 16L to 8D. It is shown that the photoperiod has a determining role in the induction of chrysopa perla prepupa diapause.
TABLE 2 diapause induction rate of chrysopa perla under different temperature and photoperiod combination conditions
Note: data in the table are mean ± sem, with different lower case letters on the same column indicating significant difference (P < 0.05).
(2) Effect of temperature on diapause Induction
As shown in tables 2 and 3, temperature also had a significant effect on diapause induction of chrysopa perla prepupa, but the effect was lower than that of photoperiod; the temperature has a regulating effect on diapause induced photoperiod reaction.
Under the same photoperiod condition, the diapause rate of prepupa is slightly reduced along with the temperature rise. Particularly under the condition of 8L to 16D short illumination, the diapause rate is obviously reduced along with the increase of temperature, but the overall reduction range is not large, and the diapause rate is higher than 90 percent. The highest diapause rate is 100 percent when the temperature is 18 ℃ and 20 ℃; when the temperature is increased to 22 ℃ and 24 ℃, the diapause rate is obviously reduced and is respectively 96.7 percent and 93.2 percent; at a temperature of 26 ℃, the diapause rate is reduced to 91.1 percent, which is obviously lower than that at 18 ℃ and 20 ℃, but the difference between the diapause rate and that at 22 ℃ and 24 ℃ is not obvious. The low temperature can obviously improve the diapause rate of the prepupa, and the temperature has a certain regulating effect on the prepupa diapause induced by photoperiod.
TABLE 3 influence of different temperatures on chrysopa perla diapause induction under 8L:16D short-light illumination
| Temperature (. degree.C.) | Observing cocoon number (head) | Diapause rate (%) | Days required for egg-cocooning |
| 18 | 111 | 100a | 28-35 days |
| 20 | 101 | 100a | 23-28 days |
| 22 | 119 | 96.7±0.7b | 19-24 days |
| 24 | 117 | 93.2±0.6b | 15-20 days |
| 26 | 192 | 91.1±3.3b | 13-16 days |
Note: data in the table are mean ± sem, with different lower case letters on the same column indicating significant difference (P < 0.05).
(3) Effect of photoperiod and temperature on diapause Induction
The results of two-factor ANOVA of diapause rate under different photoperiods and temperatures (18 ℃, 20 ℃ and 26 ℃) indicate that not only photoperiods (F)(4,30)＝724.5，df＝4,P<0.001) and temperature (F)(2,30)＝47.53，df＝2,P<0.001) has significant effect on diapause induction of chrysopa perla and significant interaction of photoperiod and temperature (F)(8,30)＝17.31，df＝8, P<0.001). The pre-pupa has 100% of diapause rate at 18 ℃ and 8L:16D under the temperature and light combination condition, and the diapause rate at 26 ℃ and 8L:16D under the temperature and light combination condition can still reach 91.1%. It is shown that although low temperature has a promoting effect on the diapause induction of prepupa, the photoperiod has a leading effect on the occurrence of prepupa diapause, and the temperature has a certain regulating effect along with the photoperiod.
The result of the comprehensive experiment 2 shows that the photoperiod is the main environmental factor for determining diapause of chrysopa perla, and more than 90% of prepupa can be induced to enter diapause by short illumination every day for less than 12 hours; the temperature has a certain regulating effect on diapause of the chrysopa perla, the diapause can be further promoted by low temperature, but the influence effect of the temperature is lower than that of the photoperiod. The whole larval stage of the chrysopa perla is raised under the conditions of 8L of short illumination at a temperature of 16D and at a temperature of 18 ℃ or 20 ℃ until the chrysopa perla is cocooned, and then 100% of individuals can be induced to enter a diapause state. In practical production application, considering economic cost factors, because the larvae develop slowly at 18 ℃ and the time required for inducing the larvae to enter diapause is longer (table 3), 20 ℃ and short illumination of 8L to 16D are superior combined conditions for inducing the larvae of the chrysopa perla to diapause, so that the prepupa can be induced to enter the diapause in a relatively short time, 100% of diapause individuals can be obtained, and the practical significance in production is achieved.
Diapause maintenance is the premise and the basis for prolonging the storage period of natural enemy insect products, and is also a key link for combining development regulation and production of natural enemy insects. In order to determine the method for diapause maintenance and storage of chrysopa perla, the applicant conducted the following experiments, and the individual steps of the method were verified from the following experiments.
Experiment 3 diapause relief test for chrysopa perla
3.1 insect source for test and feeding method: as in experiment 1 above.
3.2 Experimental methods
As is clear from the results of experiment 2, the rate of diapause induction of chrysopa perla was the highest and reached 100% at 20 ℃ under 8L 16D. Therefore, the chrysopa perla diapause prepupa (10 days after cocooning) induced under the condition is selected as a test object in the test. The diapause prepupa is respectively treated as follows: 1) directly placing at 26 deg.C and 16L:8D (control group) for continuous observation; 2) directly placing at 20 deg.C and 8L at 16D for continuous observation; 3) treating at 0 deg.C under full darkness for 10, 30, and 60 days, and observing at 26 deg.C under 16L 8D; 4) treating at 5 deg.C under full darkness for 30, 60, and 90 days, and observing at 26 deg.C under 16L 8D. The treated chrysopa perla is observed day by day, the eclosion time, the eclosion quantity and the like of the adults are recorded, and the time length and the eclosion rate from the cocoon transferring day to the adult eclosion are counted. If the diapause cocoon can normally emerge, the diapause releasing individual is determined, and the time from the cocoon transferring day to the adult emergence is used as the diapause releasing period. More than 60 chrysopa perla cocoons are observed in each treatment.
3.3 data analysis
All data in the experiment were analyzed statistically and the results are expressed as mean ± sem. The comparison of the emergence rate adopts chi-square test; reciprocal transformation is carried out at the diapause release stage, then One-Way ANOVA in GraphPad Prism V6.03 software is used for variance analysis, and Tukey multiple comparison method is adopted for difference significance test.
3.4 analysis of results
The effect of different temperature and illumination combination conditions on the diapause release of chrysopa perla is shown in Table 4, and it is known from the table that diapause prepupa can release diapause under high-temperature long illumination conditions (26 ℃,16L:8D) and medium-temperature short illumination conditions (20 ℃,8L:16D), but the time required for diapause release is longer, the diapause release rate is lower, and the eclosion of adults is irregular. Under the conditions of temperature and light combination of 26 ℃ and 16L:8D, the diapause release period takes 72 days on average, and about 50 percent of individuals can release diapause and eclose into adults. Under the conditions of 20 ℃ and 8L at the speed of 16D, although the diapause release rate is relatively high, the time required from the prepupa diapause release to the eclosion of the imagoes is obviously prolonged, the average time is 107 days, the eclosion duration period of the imagoes is obviously prolonged, and the rapid diapause release is not facilitated.
TABLE 4 influence of different temperature and light combination conditions on diapause relief of chrysopa perla
| Combined conditions of temperature and light | Diapause release rate (%) | Average diapause release calendar (day) | Number of days of eclosion of adult |
| 26℃，16L:8D | 49.1a | 71.9±20.8a | 40-108 |
| 20℃，8L:16D | 86.7b | 106.8±20.9b | 83-186 |
The effect of low temperature on the diapause release of chrysopa perla is shown in tables 4 and 5, and the results show that the zero-temperature low temperature is not beneficial to the survival and diapause release of chrysopa perla, only few chrysopa perla can release diapause and restore growth and development after being treated at 0 ℃ for a period of time and then transferred to proper conditions, the diapause release rate is extremely low, and the time required for diapause release is long.
TABLE 5 Effect of Low temperature 0 ℃ on the diapause relief of chrysopa perla
| Days of treatment at 0 ℃ | Diapause release rate (%) | Average diapause release calendar (Tian) | Number of days of eclosion of adult |
| 10 | 8.4b | 98.8±29.3a | 61-160 |
| 30 | 30.1a | 53.2±28.0b | 27-110 |
| 60 | 16.2a | 36.8±7.2b | 22-48 |
After short-term treatment at moderate low temperature (5 ℃), the chrysopa septempunctata is transferred to normal development conditions to obviously promote the diapause relieving process of chrysopa septempunctata. The results in Table 6 show that the diapause pupae were treated at 5 ℃ for 30 days, the diapause release rate was less than 60%, and 34.8 days were required on average from diapause release to adult eclosion. After the treatment lasts for more than 60 days, the diapause removal rate is obviously increased and can reach more than 85 percent, and the maximum rate is 94 percent, which is obviously higher than that of other treatment groups; the diapause release period is also obviously shortened, and most of the diapause release period can emerge into adults within 21-23 days; the eclosion uniformity of the imagoes is also obviously improved, and the imagoes can be concentrated within 15 days to complete the eclosion. The results show that the chrysopa perla prepupa is required to be subjected to certain low-temperature stimulation when the diapause is removed, but when the low-temperature treatment time is too short, the diapause cannot be completely removed, so that part of individuals cannot recover growth and development under proper conditions; only after the low-temperature treatment time exceeds a certain range (treatment at 5 ℃ for more than 60 days), can most of chrysopa perla relieve diapause and emerge into adults in a short period.
TABLE 6 influence of Low temperature 5 ℃ on the diapause relief of chrysopa perla
| Days of treatment at 5 ℃ | Diapause release rate (%) | Average diapause release calendar (Tian) | Number of days of eclosion of adult |
| 30 | 59.5b | 34.8±11.2a | 19-69 |
| 60 | 93.5a | 23.1±3.9b | 17-35 |
| 90 | 85.5a | 21.6±2.3b | 18-30 |
According to the results of the comprehensive experiment 3, the delayed diapause release progress of the chrysopa perla can be remarkably promoted by moderate low-temperature treatment for a certain time, from the perspective of practical production and application, the prepupa in the delayed state is treated under the conditions of 5 ℃, full darkness and 60 days by combining factors such as delayed diapause release rate, delayed diapause release duration, adult eclosion uniformity, vitality and the like, then transferred to normal development conditions (26 ℃,16L:8D), more than 94% of individuals can be subjected to delayed diapause release within 23 days on average, eclosion is converted into adults, normal predation, mating, oviposition, reproduction and the like are carried out, and the pest control efficiency of natural enemy insect products is exerted.
While one embodiment of the present invention has been described in detail, the description is only a preferred embodiment of the present invention and should not be taken as limiting the scope of the invention. All equivalent changes and modifications made within the scope of the present invention shall fall within the scope of the present invention.
Claims (5)
1. A diapause relieving technology for chrysopa perla is characterized in that: the method comprises the following steps:
1) selecting chrysopa perla prepupa entering a diapause state as a diapause removing object;
2) placing the chrysopa perla prepupa selected in the step 1) in a low-temperature environment for short-term treatment, wherein the low-temperature environment condition is that the photoperiod is 0L to 24D, the temperature is 5 ℃, and the continuous treatment is carried out for more than or equal to 60 days;
3) transferring the chrysopa perla prepupa treated in the step 2) to a condition that the temperature is 26 ℃ and the photoperiod is 16L:8D, so that the chrysopa perla is released from diapause and eclosion into adults.
2. The diapause release technique for chrysopa perla according to claim 1, characterized in that: the chrysopa perla diapause prepupa selected in the step 1) is a cocoon of 10 days old after cocooning.
3. The diapause release technique for chrysopa perla according to claim 1, characterized in that: the diapause release is performed in an artificial climate box.
4. The diapause release technique for chrysopa perla according to claim 1, characterized in that: the diapause release rate of the chrysopa perla prepupa in the step 3) is 90-95%, and the average eclosion period of the adults is 23 days.
5. The diapause release technique for chrysopa perla according to claim 1, characterized in that: the step 2) is continued for 60 days.
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