CN108207818B - Diapause maintaining and storing method for chrysopa perla - Google Patents
Diapause maintaining and storing method for chrysopa perla Download PDFInfo
- Publication number
- CN108207818B CN108207818B CN201810048666.2A CN201810048666A CN108207818B CN 108207818 B CN108207818 B CN 108207818B CN 201810048666 A CN201810048666 A CN 201810048666A CN 108207818 B CN108207818 B CN 108207818B
- Authority
- CN
- China
- Prior art keywords
- diapause
- chrysopa
- chrysopa perla
- perla
- temperature
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000005058 diapause Effects 0.000 title claims abstract description 179
- 241000021603 Chrysopa perla Species 0.000 title claims abstract description 102
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000005286 illumination Methods 0.000 claims abstract description 34
- 238000003860 storage Methods 0.000 claims abstract description 29
- 238000012423 maintenance Methods 0.000 claims abstract description 21
- 230000003203 everyday effect Effects 0.000 claims abstract description 8
- 230000006698 induction Effects 0.000 claims description 24
- 230000032669 eclosion Effects 0.000 claims description 17
- 230000019617 pupation Effects 0.000 claims description 7
- 230000007774 longterm Effects 0.000 claims description 5
- 230000001418 larval effect Effects 0.000 claims description 4
- 241001064674 Chrysopa septempunctata Species 0.000 claims description 3
- 230000008030 elimination Effects 0.000 claims 1
- 238000003379 elimination reaction Methods 0.000 claims 1
- 241000607479 Yersinia pestis Species 0.000 abstract description 19
- 238000011282 treatment Methods 0.000 abstract description 12
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 230000002265 prevention Effects 0.000 abstract description 5
- 230000009471 action Effects 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 241000238631 Hexapoda Species 0.000 description 48
- 238000011161 development Methods 0.000 description 41
- 230000018109 developmental process Effects 0.000 description 41
- 238000002474 experimental method Methods 0.000 description 27
- 230000000694 effects Effects 0.000 description 19
- 235000013601 eggs Nutrition 0.000 description 17
- 238000005516 engineering process Methods 0.000 description 10
- 230000002035 prolonged effect Effects 0.000 description 10
- 230000001105 regulatory effect Effects 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 241000382353 Pupa Species 0.000 description 6
- 230000001276 controlling effect Effects 0.000 description 6
- 230000008901 benefit Effects 0.000 description 5
- 230000033228 biological regulation Effects 0.000 description 5
- 230000001939 inductive effect Effects 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 241001124076 Aphididae Species 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 4
- 241001292007 Chrysopa Species 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000011217 control strategy Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 241000123346 Chrysosporium Species 0.000 description 2
- 241000258937 Hemiptera Species 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000011419 induction treatment Methods 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 241000238876 Acari Species 0.000 description 1
- 241000253994 Acyrthosiphon pisum Species 0.000 description 1
- 241001026766 Chrysopa formosa Species 0.000 description 1
- 241000258935 Chrysopidae Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000255777 Lepidoptera Species 0.000 description 1
- 241000594036 Liriomyza Species 0.000 description 1
- 241000171274 Megoura Species 0.000 description 1
- 241001537210 Perna Species 0.000 description 1
- 241000255129 Phlebotominae Species 0.000 description 1
- 244000184734 Pyrus japonica Species 0.000 description 1
- 240000003461 Setaria viridis Species 0.000 description 1
- 235000002248 Setaria viridis Nutrition 0.000 description 1
- 241001414989 Thysanoptera Species 0.000 description 1
- 240000006677 Vicia faba Species 0.000 description 1
- 235000010749 Vicia faba Nutrition 0.000 description 1
- 235000002098 Vicia faba var. major Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000000546 chi-square test Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000009025 developmental regulation Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011328 necessary treatment Methods 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides a diapause maintaining and storing method of chrysopa perla, which comprises the following steps: 1) selecting the chrysopa perla entering the diapause state as a diapause maintaining object, wherein the chrysopa perla in the diapause state is in a prepupuncture period; 2) placing the chrysopa perla selected in the step 1) in a diapause maintaining environment, and controlling the combination of the illumination time and the temperature every day to maintain the chrysopa perla in a diapause state; the illumination time length of each day adopts a light cycle (0-10) L (14-24) D, the temperature is (0-10) DEG C, and the storage time length is 30-300 days. The beneficial effects of the invention are: the method realizes the accurate control of the diapause maintenance of the chrysopa perla, prolongs the shelf life of the product and the prevention and control action time in the field, ensures the annual large-scale propagation production, provides sufficient natural enemy products for the biological control of various agricultural and forestry pests, and improves the scientific treatment level of the agricultural pests.
Description
Technical Field
The invention relates to the field of biotechnology, in particular to a diapause maintaining and storing method of chrysopa perla, which can maintain the chrysopa perla pupa which enters diapause in a development stasis state for a certain time by a diapause maintaining technology so as to achieve the purpose of prolonging the storage period of the chrysopa perla.
Background
In recent years, with the change of modern agricultural farming modes and the comprehensive result of global climate change, the agricultural pest population structure, the catastrophe rule 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 prevent and 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 maintaining and storing method for chrysopa perla, which realizes the accurate control of the development progress of a good predatory natural enemy insect chrysopa perla 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 method for diapause maintenance and storage of chrysopa perla, comprising the steps of:
1) selecting the chrysopa perla entering the diapause state as a diapause maintaining object, wherein the chrysopa perla in the diapause state is in a prepupuncture period;
2) placing the chrysopa perla selected in the step 1) in a diapause maintaining environment, and controlling the combination of the illumination time and the temperature every day to maintain the chrysopa perla in a diapause state; the illumination time length of each day adopts a light cycle (0-10) L (14-24) D, the temperature is (0-10) DEG C, and the storage time length is 30-300 days.
Further, the photoperiod in the step 2) is 0L to 24D, the temperature is 5 ℃, and the storage time is 300 days.
Further, the pre-pupation period in the step 1) is cocoons which are 10 days old after cocoons are formed.
Further, the diapause maintenance and long-term storage are carried out in a climatic chamber.
Furthermore, after the diapause maintenance period is finished, 85-95% of individuals can release diapause and eclose into adults within 15-25 days on average under the conditions of 26 ℃ and 16L: 8D.
The invention has the beneficial effects that:
the diapause littlefly is stored under the condition of full darkness at 5 ℃, the storage period can be maintained for 30-300 days, more than 85 percent of individuals can normally emerge into adults after the diapause of the littlefly stored by the method is removed, and normal predation, mating, oviposition, reproduction and the like are carried out, so that the pest control efficiency of natural enemy insect products is exerted. By utilizing the technology, the chrysopa perla can be produced all the year round, and the product is stored by the diapause technology, so that sufficient natural enemy products are provided for the biological control of various agricultural and forestry pests, the control action time of the natural enemy products is prolonged, the biological control means is enriched, and the scientific control level of the agricultural pests is improved.
Drawings
FIG. 1 shows the diapause maintenance time and survival rate of chrysopa perla under the conditions of 26 ℃ and 16L: 8D;
FIG. 2 is diapause maintenance time and adult emergence rate of chrysopa perla at low temperature 0 ℃ (different lower case letters indicate significance of difference P < 0.05);
FIG. 3 is diapause maintenance time and adult emergence rate of chrysopa perla at low temperature of 5 ℃ (different lower case letters indicate significance of difference P < 0.05).
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 pea Setaria viridis under the conditions of temperature of 26 +/-1 ℃ and photoperiod of 16L:8D (artificial climate box, Ningbo Jiangnan RXZ-500B), collecting Chrysopa perla eggs by the same method, and adding a sufficient amount of aphids every day for feeding after the eggs hatch larvae. 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 un-eclosion cocoons are dissected, the survivors are still maintained, namely, the calluses are considered as diapause, and the survival insect state is diapause insect state. 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 chrysopa perla is diapaused in the cocoon in the prepupa state, and once it enters diapause, it does not pupate even if it is 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. Diapause induction treatment is only carried out on different development stages before the 3 rd larva and development insect states after the 3 rd larva, no diapause individual appears, all chrysopa cochleri can normally pupate, eclose and lay eggs, and the result is consistent with the result that all development stages are under normal development Conditions (CK), and shows that the larva at the egg stage, the larva at the 1 st age, the larva at the 2 nd age, the larva at the 1-2 nd age and the prepupa are insensitive 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 climatic 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 the 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 produce diapause, and the diapause rate is 0.
The diapause induction law of photoperiod is the same as that at 18 ℃ at 20 ℃. The diapause rate is the highest and reaches 100 percent 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 remarkably reduced, and no diapause individual appears under the condition of long illumination for 16 h.
According to the photoperiod response curves of chrysopa perla at 18, 20 and 26 ℃, the critical photoperiod of the chrysopa perla at each temperature is 14.4,14.3 and 12.9h respectively by calculating the relation between the illumination duration and the diapause rate, and the critical photoperiod of the chrysopa perla gradually shortens with the increase of the temperature (figure 1).
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, reaching 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 short illumination of 8L:16D, 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 short illumination 8L 16D conditions different temperature on the effect of chrysopa perla diapause induction
| Temperature (. degree.C.) | Observing cocoon number (head) | Diapause rate (%) | Days required for egg-cocooning |
| 18 | 111 | 100a | 28-35 |
| 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 incubation rate of prepupa is 100% under the combined condition of temperature and light at 18 ℃ and 8L:16D, and the incubation rate can still reach 91.1% under the combined condition of temperature and light at 26 ℃ and 8L: 16D. 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 is that the photoperiod is the main environmental factor for determining diapause of the chrysopa perla, and more than 90 percent 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 maintenance and storage test of chrysopa perla
3.1 insect source to be tested and raising method
The insect source and the feeding method for the experiment are as described in experiment 1.
3.2 Experimental methods
As is clear from the results of the above experiment 1, 100% of chrysopa perla individuals were induced to enter diapause under the combined conditions of temperature and light at 20 ℃ and 8L:16D, and therefore all the prepupoles (cocoons, 10 days after cocooning) of diapause chrysopa perla used in this experiment were induced under the conditions of 20 ℃ and 8L: 16D. The specific steps and method of the diapause maintaining experiment are as follows: respectively storing the diapause prepupa (in a cocoon shell) under the following conditions: 1) conditions (control CK) at 26 ℃ and 16L: 8D; 2) at 0 ℃, under the condition of full darkness; 3)5 ℃ and in the dark. Taking out a batch of cocoons at 0 ℃ every 10 days and at 5 ℃ every 30 days, transferring the cocoons to 26 ℃,16L:8D for continuous development, observing and recording the eclosion time, quantity and the like of adults under various treatment conditions day by day, and counting the time length and the eclosion rate from the cocoon transferring day to the adult emergence. If the diapause cocoon can normally emerge, it is regarded as the diapause-released individual. More than 60 chrysopa perla calluses are observed in each treatment.
Control experiment of normal development group of chrysopa septempunctata: the normal development prepupa (in the cocoon shell) of chrysopa perla bred at 26 ℃ under 16L:8D were subjected to the following conditions: 1) at 0 ℃, under the condition of full darkness; 2) at 5 ℃, in a dark condition; 3)10 ℃ and under the condition of full darkness. Taking a batch of cocoons under each treatment condition every 10 days, transferring the cocoons to a container with the temperature of 26 ℃, the volume of 16L:8D for continuous development, observing and recording the eclosion time, the number and the like of adults day by day, and counting the time length and the eclosion rate required from the day of transferring cocoons to the eclosion of adults.
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 and utilizes SPSS 20.0 to analyze; the time length required by the eclosion of the imagoes is subjected to reciprocal transformation, 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
26 ℃,16L: under the condition of 8D, the chrysopidae diapause cocoons can maintain diapause for 40-108 days, the average diapause maintenance period (storage period) is 71.9 days, but only 49.1 percent of individuals can relieve diapause, recover growth and develop and emerge into adults, and the emergence rate is low, as shown in figure 1.
The influence of the low temperature of 0 ℃ on the diapause maintenance of the chrysopa perla prepupa is shown in figure 2, and the figure shows that the low temperature of 0 ℃ is not favorable for the diapause maintenance of the prepupa, the chrysopa perla prepupa is stored for 10-60 days and then transferred to the normal development condition (26 ℃,16L:8D) for continuous development, compared with the control group (26 ℃,16L:8D), the diapause prepupa has high death rate, the rate of the diapause prepupa which can be released for continuous development to be imagoes is lower, and the eclosion rate is only 30% at most. It is proved that the low temperature of 0 ℃ is not favorable for maintaining diapause, and the death rate of chrysopa perla prepupa at the temperature is higher, so that the chrysopa perla prepupa is not suitable for long-term storage of diapause cocoons.
As shown in FIG. 3, the diapause maintenance of chrysopa perla prepupa can be significantly promoted at a low temperature of 5 ℃. When the diapause chrysopa perla prepupa is stored for 30 days at the temperature of 5 ℃, the eclosion rate is 58.2 percent and is higher than that of a control group, but the difference between the eclosion rate and the eclosion rate is not obvious. When the compound fertilizer is stored for 60 to 300 days, after the compound fertilizer is transferred to a place with the temperature of 26 ℃ and 16L:8D for diapause removal, the eclosion rate can reach more than 85 percent and can reach 93.3 percent at most, and the eclosion rate is not obviously different among treatments but is obviously higher than that of a control group and a 30-day treatment group. In addition, as the storage time is prolonged, the emergence uniformity of the chrysopa perla prepupa imago after diapause release is gradually improved, and the time required by emergence is obviously shortened. It is demonstrated that the 5 ℃ all-black condition is helpful for diapause maintenance and long-term storage of chrysopa perla prepupa.
In said fig. 2-3, CK: the diapause release rate is at 26 ℃ and 16L: 8D; different lower case letters represent significance of difference P < 0.05.
The results of the experiment in the normal development group are shown in Table 4, and all individuals died when the chrysopa perla normal development preputial was treated at 0 ℃ and 5 ℃ for 10, 20, and 30 days, respectively. The pre-pupa emergence rate can reach more than 60% after being stored for 10, 20 and 30 days at 10 ℃, and the emergence rate of the pre-pupa in normal development is not obviously different among treatments, but is obviously reduced in a treatment group for 30 days compared with a control group. Along with the extension of the low-temperature storage time, the time required by the normal development and the individual emergence of the chrysopa septempunctata is obviously prolonged.
Compared with a normal development control group, after the diapause individuals are stored at 5 ℃ for 300 days, the eclosion rate of the diapause individuals is up to 86.4 percent under the condition of normal development, but the normal development individuals die after being stored at 5 ℃ for 10 days. Even at higher temperatures of 10 ℃, normally developing individuals can be stored for a short period of 30 days, but the emergence rate drops to 60%. Therefore, the diapause can obviously prolong the storage period of the chrysopa perla and can prolong the shelf life of chrysopa perla products by at least 10 times.
TABLE 4 Effect of storage at Low temperature of 10 ℃ for various periods of time on Normal developmental individuals of chrysopa perla
Note: data in the table are mean ± sem, with different lower case letters on the same column indicating significant difference (P < 0.05).
The result of the comprehensive experiment 3 shows that the diapause can obviously prolong the shelf life (storage period) of the product of the chrysopa perla, and the prepupa in the diapause state is stored at 5 ℃ in full darkness for 300 days. Considering from the practical production and application, by integrating the factors of emergence rate and uniformity of the emergence of imagoes, the short-term storage of the chrysopa perla can place the normally developed prepupa (newly formed cocoons) in the dark condition of 10 ℃, and the storage period is preferably no more than 30 days; the long-term storage of more than 30 days can induce the chrysopa perla to enter a diapause state, the chrysopa perla in the diapause state is stored under the condition of full darkness at 5 ℃, the storage period can be maintained at 30-300 days, more than 85 percent of individuals can normally emerge into adults after the chrysopa perla stored by the method is transferred to 26 ℃ and 16L:8D to remove the diapause, the adults can all emerge in an average range of 15-25, the emerged adults can normally prey, mate, lay eggs, reproduce and the like, 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 maintaining and storing method of chrysopa perla is characterized in that: the method comprises the following steps:
1) selecting the chrysopa perla entering the diapause state as a diapause maintaining object, wherein the chrysopa perla in the diapause state is in a prepupuncture period; wherein the diapause induction time of the chrysopa perla at least comprises 3-instar larvae in the larval stage, and the diapause induction condition of the chrysopa perla is 20 ℃, and the illumination condition is 8L to 16D;
2) placing the chrysopa perla selected in the step 1) in a diapause maintaining environment, and controlling the combination of the illumination time and the temperature every day to maintain the chrysopa perla in a diapause state; the illumination time is 0L to 24D in a photoperiod every day, and the storage is carried out for 60 to 300 days at the temperature of 5 ℃;
3) transferring the chrysopa perla in the step 2) to 26 ℃ and 16L:8D for diapause elimination, wherein the emergence rate can reach more than 85%.
2. The method for diapause maintenance and storage of chrysopa perla according to claim 1, wherein: the length of the chrysopa septempunctata in the step 2) is 300 days.
3. The method for diapause maintenance and storage of chrysopa perla according to claim 1, wherein: the pre-pupation period in the step 1) is cocoons which are 10 days old after cocoons are formed.
4. The method for the diapause maintenance and storage of chrysopa perla according to claim 1, wherein: after the prepupa in the step 1) is subjected to the diapause maintenance period, 85-95% of individuals can be transferred to the conditions of 26 ℃ and 16L:8D to release the diapause in 15-25 days on average, and eclosion is realized to be imagoes.
5. The method for diapause maintenance and storage of chrysopa perla according to claim 1, wherein: the diapause maintenance and long-term storage are carried out in an artificial climate box.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810048666.2A CN108207818B (en) | 2018-01-18 | 2018-01-18 | Diapause maintaining and storing method for chrysopa perla |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810048666.2A CN108207818B (en) | 2018-01-18 | 2018-01-18 | Diapause maintaining and storing method for chrysopa perla |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108207818A CN108207818A (en) | 2018-06-29 |
| CN108207818B true CN108207818B (en) | 2022-05-24 |
Family
ID=62667251
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810048666.2A Active CN108207818B (en) | 2018-01-18 | 2018-01-18 | Diapause maintaining and storing method for chrysopa perla |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108207818B (en) |
-
2018
- 2018-01-18 CN CN201810048666.2A patent/CN108207818B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108207818A (en) | 2018-06-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | The biology and ecology of the emerald ash borer, Agrilus planipennis, in China | |
| Niemelä et al. | The effect of previous damage on the quality of Scots pine foliage as food for diprionid sawflies | |
| CN102573453B (en) | Method for producing viable tetraploid oysters | |
| Gols et al. | Temporal changes affect plant chemistry and tritrophic interactions | |
| Choochote et al. | Systematic techniques for the recognition of Anopheles species complexes | |
| Zheng et al. | Development and reproduction of the hoverfly Eupeodes corollae (Diptera: Syrphidae) | |
| Oloo | The role of local natural enemies in population dynamics of Chilo partellus (Swinh.)(Pyralidae) under subsistence farming systems in Kenya | |
| Mathenge et al. | The biology of Dactylopius tomentosus (Hemiptera: dactylopiidae) | |
| Cock | The use of parasitoids for augmentative biological control of pests in the People's Republic of China. | |
| Maroufpoor et al. | Life table parameters of Neoseiulus californicus (Acari: Phytoseiidae), on the European red mite, Panonychus ulmi (Acari: Tetranychidae) in laboratory condition | |
| CN105165726A (en) | Method for realizing three-dimensional propagation of Aphidius gifuensis Ashmaed by means of cuttage plant | |
| CN102771447B (en) | Diapause maintaining method of Coccinella septempunctata | |
| CN108207818B (en) | Diapause maintaining and storing method for chrysopa perla | |
| CN108077192B (en) | Diapause relieving technology for chrysopa perla | |
| Bashir et al. | Observations on the effect of the weevils Neochetina eichhorniae Warner and Neochetina bruchi Hustache on the growth of water hyacinth | |
| CN108056071A (en) | A kind of Diapause inducement method of beautiful Chrysopa | |
| Gorder et al. | Life history of the parsnip webworm, Depressaria pastinacella (Lepidoptera: Oecophoridae), in central Iowa | |
| Sultana et al. | Mating behaviour of Hieroglyphus species (Hemiacridinae: Acrididae: Orthoptera) from Pakistan | |
| Amevoin et al. | Why Callosobruchus rhodesianus causes limited damage during storage of cowpea seeds in a tropical humid zone in Togo | |
| Thongwat et al. | Crossing experiments supporting the specific status of Anopheles maculatus chromosomal form K | |
| Wilson et al. | The introduction into Australia of parasites of Listroderes obliquus Klug | |
| Eltayba et al. | Relation between female cocoons (pupae) measurements and the number of eggs laid (fecundity) by the wild silk moth Epiphora bauhiniae (Gurin-meneville) Lepidopetera: saturniidae | |
| CN110367207B (en) | Method for breeding large amount of trichogramma matsutake with convex legs | |
| Yukawa et al. | Improvement in the practical use of an aphidophagous gall midge, Aphidoletes aphidimyza (Diptera: Cecidomyiidae), in greenhouses. | |
| CN109997794B (en) | Application of sulfadiazine in reproduction behavior of Wolbachia infected short-tube trichogramma |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |