CN104351140A - Leis dimidiata diapause relieving method - Google Patents
Leis dimidiata diapause relieving method Download PDFInfo
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- CN104351140A CN104351140A CN201410495765.7A CN201410495765A CN104351140A CN 104351140 A CN104351140 A CN 104351140A CN 201410495765 A CN201410495765 A CN 201410495765A CN 104351140 A CN104351140 A CN 104351140A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Abstract
The invention discloses a leis dimidiata diapause relieving method which comprises the following steps: 1) selecting leis dimidiata imago in the diapause condition as diapause relieving objects; 2) placing the leis dimidiata selected in the step 1) under the diapause relieving environment, and through regulation and control of combination of everyday lighting duration, temperature and total lighting duration, enabling the leis dimidiata to lay eggs so as to relieve diapause, wherein the everyday lighting duration adopts photoperiod (8-14)L:(16-10)D, the temperature is 24 DEG C, and the treatment continues for 72 h. Through the control of environmental factor and the combination of necessary treatment measures, the accurate control on growth start time of an excellent predatory natural enemy insect, namely the leis dimidiata, is realized, the diapause relieving rate can reach more than 98%, and technical support is provided for annual mass production of the leis dimidiata.
Description
Technical field
The present invention relates to a kind of red shoulder ladybug Diapause termination method, can will be recovered vitality by the red shoulder grandis adults of diapause by the method, comprise fecundity, prey ability, circle in the air and expand numerous ability, guarantee that natural enemy product plays the usefulness of Control pests.
Background technology
The crop yield loss that China causes because of agricultural pest is every year 10-15%, realizes agricultural pest science and administers, and for the strategy implementation of guarantee national food security, promotes that agricultural high yield has important economical, societal benefits.Aphid is the first insect of agricultural production, and the whole world is every year because of the loss nearly 5,000,000,000 dollars that aphid is caused, and the current chemical insecticide control aphid class that mainly depends on endangers.
In control pest process, if depend on chemical pesticide simply, the negative issue of the aspects such as a series of ecology, environment, food security will be brought.Due to the Long-Time Service of chemical pesticide, some insects have produced very strong pesticide resistance, and the natural enemy of various pests is killed in a large number, cause some Pest rampancies to endanger.In addition, chemical pesticide meeting polluted-water, air and soil, and be enriched to human body by food chain, harm population health.Utilize biotechnology to carry out prevention and control damage by disease and insect, just can effectively avoid above-mentioned shortcoming, thus there is wide development prospect.Natural enemy insect is wherein utilized to be exactly one of effective biological control means.
A bottleneck problem of the outer natural enemy insect industrialization of Present Domestic is: it is longer that natural enemy insect expands numerous cycle, survival period is shorter, the product export time is difficult to consistent with Injurious insect emergence period, often causes natural enemy with regard to dead before insect occurs, or does not have natural enemy product to use when insect occurs.So the development progress of regulation and control natural enemy insect product has important practical significance.Realize the regulation and control to natural enemy insect product development progress, just can utilize insect heritability inherently---diapause.
Red shoulder ladybug Leis dimidiata (Fabricius) belongs to coleoptera ladybug, come from Palaearctic region, nowadays all distribution is had in the greater part, the world, the ground such as Beijing, Jilin, Liaoning, the Inner Mongol, Hebei, Shandong, Henan, Sichuan, Ningxia, Yunnan and Tibet are mainly distributed in China, the multiple aphid (Jing Ying etc., 2002) on crops and fruit tree can be taken food.Zong Liang Ping etc. (1988) research finds that red shoulder ladybug likes best predation cotten aphid Aphis gossypii Glover, but also preys on multiple aphids such as various wheat aphid, sorghum aphid, soybean aphid Aphiscraccivora Koch, acacia aphid Aphis robiniae Macchiati.Ladybug is eating aphid generally, but red shoulder ladybug is when aphid lacks, and also takes food tetranychid (Wang Yunhua etc., 1984).Red shoulder ladybug is as one of the Dominant Natural Enemies of aphid, and its population quantity is large, and reproduction speed is fast, studying its predation functional responses and is intended to scientific evaluation natural enemy control action, providing scientific basis for utilizing nature enemy in amount better.
In recent years, red shoulder ladybug by development and application in succession, and achieves significant ecological benefits and economic benefit in many countries, has played irreplaceable prevention and control effect in the biological control of aphid.But in production application, normal exist that to expand numerous cycle short, product can not long storage periods and can not apply in good time, imaginal diapause and affect problems such as expanding numerous efficiency, become the technical bottleneck of restriction red shoulder ladybug large-scale production application.
Summary of the invention
For problems of the prior art, the invention provides a kind of red shoulder ladybug Diapause termination method, by manipulation envirment factor, as every day light irradiation time, temperature etc., in conjunction with necessary treatment measures, realizing excellent predatory natural enemy insect---red shoulder ladybug grows the accurate control of time started.
The object of the invention is to be achieved through the following technical solutions:
Red shoulder ladybug Diapause termination method, the method comprises the following steps:
1) choose enter diapause status red shoulder grandis adults as Diapause termination object;
2) by step 1) under the red shoulder ladybug chosen is placed in Diapause termination environment, by the combination of regulation and control light irradiation time every day, temperature, total light irradiation time three, make red shoulder ladybug start to lay eggs thus termination of diapause; Described light irradiation time employing every day photoperiod (8 ~ 14) L:(16 ~ 10) D, temperature is 24 DEG C, continues process 72h.Here 72h refers to lasting process 3 days.
Further, described step 2) middle light irradiation time employing every day photoperiod 10L:14D.
Further, described step 2) in also to regulate and control the humidity of diapause maintenance environment, relative moisture is 70 ± 10%.
Further, described step 1) in the red shoulder ladybug chosen namely enter diapause status in the first emerging adult stage, first emerging adult is the adult within just sprouting wings 12 hours.
Further, described Diapause termination carries out in climatic cabinate.
Further, after described red shoulder ladybug enters diapause status, through the storage duration of 300 ~ 400d.
The present invention is to enter the red shoulder ladybug of diapause status as Diapause termination object, to the time of diapause maintenance particular/special requirement useless, if use the red shoulder grandis adults of diapause of long storage periods as releasing object, then can adopt following red shoulder ladybug diapause maintenance and long storage periods method.
Red shoulder ladybug diapause maintenance and long storage periods method, the method comprises the following steps:
1) choose enter diapause status red shoulder ladybug as diapause maintenance object, namely the red shoulder ladybug of this state enters diapause status in the first emerging adult stage;
2) by step 1) under the red shoulder ladybug chosen is placed in diapause maintenance environment, by the combination of regulation and control light irradiation time every day, both temperature, make red shoulder ladybug maintenance diapause status; Described light irradiation time employing every day photoperiod (2 ~ 6) L:(22 ~ 18) D, temperature is 10 ~ 15 DEG C, and storage duration is 300 ~ 400d.
Further, described step 2) middle light irradiation time employing every day photoperiod 2L:22D, temperature is 13 DEG C, storage duration >=365d.
Further, described step 2) in also to regulate and control the humidity of diapause maintenance environment, relative moisture is 70 ± 10%.
Further, described step 1) in just emerging adult be the adult within just sprouting wings 12 hours.
Further, described diapause maintenance and long storage periods carry out in climatic cabinate.
The red shoulder ladybug entering diapause status can be obtained by following Diapause inducement method.
Red shoulder ladybug Diapause inducement method, the method comprises the following steps:
1) determine that the sensitive period that the diapause of red shoulder ladybug regulates and controls is first emerging adult, choose the red shoulder ladybug of this worm state as Diapause inducement object;
2) by step 1) under the red shoulder ladybug chosen is placed in Diapause inducement environment, by the combination of regulation and control light irradiation time every day, temperature, total induction duration three, make red shoulder ladybug enter diapause status; Described every day, light irradiation time adopted photoperiod (8 ~ 10) L:(16 ~ 14) D, temperature is 13 ~ 15 DEG C, successive induction time >=192h.Here 192h refers to 8 days.
Further, described step 2) in every day light irradiation time be 8hr/d, temperature is 13 DEG C, and total induction duration is 192h.
Further, described step 2) in also to regulate and control the humidity of Diapause inducement environment, relative moisture is 70 ± 10%.
Further, described step 2) in the successive induction time be 240h or 288h.
Further, described step 1) in just emerging adult be the adult within just sprouting wings 12 hours.
Further, described Diapause inducement carries out in climatic cabinate.
Further, before choosing Diapause inducement object, take on four-age larva, the mature larva of ladybug to red and all implement short photoperiod process pupa time, every day, light irradiation time was 8 ~ 10hr/d.
The red shoulder ladybug Diapause termination method of above-mentioned technical characteristic is adopted to have the following advantages:
The present invention is directed to red shoulder ladybug Leis dimidiata (Fabricius) this excellent predatory natural enemy insect, carry out and utilized the invention of this worm development progress of the cytokine regulatory such as photoperiod, thermoperiod to study, carry out system research for Diapause termination, achieve the control accurate to red shoulder ladybug development progress.Carry out the red shoulder ladybug stored with diapause status, its product shelf phase never diapause within 30 days, extend to 300 days, all there are raising in various degree in the red shoulder ladybug vitality after Diapause termination such as fecundity, egg laying amount, lay eggs duration, life-span etc.Utilize this technology, can realize whole year production red shoulder ladybug, stored by diapause technology by its product, the biological control for aphid class pest provides natural enemy product, has enriched biological control means, improves the science improvement level of pest.
Carry out the diapause control technique research of red shoulder ladybug, the vast propagation of red shoulder ladybug can not only be promoted in production practices, extend its product shelf phase and the prevention and control action time in field, also development characteristics and the occurrence dynamics of grasping natural enemy insect is contributed to, improve control of insect efficiency, also contribute to deepening the understanding to natural enemy insect developmental mechanism, explore insect to the adaptation mechanism of environment and evolutionary path.
Accompanying drawing explanation
Fig. 1 is the ovary displaing micro picture of the female worm of diapause red shoulder ladybug;
Fig. 2 is the ovary canal displaing micro picture of the female worm of diapause red shoulder ladybug;
Fig. 3 is the ovary displaing micro picture of the female worm of non-diapause red shoulder ladybug;
Fig. 4 is the ovary canal displaing micro picture of the female worm of non-diapause red shoulder ladybug;
Fig. 5 does temperature and photoperiod to affect statistical chart to diapause duration mutually;
When Fig. 6 is photoperiod L8:D16, temperature affects statistical chart to red shoulder ladybug Diapause termination;
When Fig. 7 is 24 DEG C, the photoperiod affects statistical chart to red shoulder ladybug Diapause termination.
Embodiment
For further setting forth the present invention for the technological means that reaches predetermined goal of the invention and take and effect, below in conjunction with accompanying drawing and preferred embodiment, structure of the present invention, workflow are described in detail as follows.
The present invention be applicant on the basis of great many of experiments, expend 5 the years cultivate, checking acquired by technological achievement.
1, time cost: from 2010 so far, lasts 5 years.
2, Financial cost:
(1) equipment: successively purchase climatic cabinate 7 (south of the River, Ningbo 500C type), expenditure funds 1.5*7=11.2 ten thousand yuan;
(2) reagent: purchase the necessary chemical reagent keeping the necessary seed of examination worm population, liquid fertilizer, vermiculite, the peat composed of rotten mosses, carry out Diapause inducement test, accumulative funds 1.5 ten thousand yuan of/year * 5 years=7.5 ten thousand yuan;
(3) artificial: except inventor works, still to employ casual labour 2 people, 1200 yuan of/month * 10 months * 5 years=60,000 yuan;
(4) place: except utilizing the laboratory of inventor unit, still rents test greenhouse, solarium, annual cost 10000 yuan, accumulative funds 50,000 yuan;
(5) consumptive material: support the test consumable such as worm frame, dependent insect cage, temperature-humidity monitoring instrument, photoperiod monitor, time switch, sunshade net, accumulative 1.5 ten thousand yuan of/year * 5 years=7.5 ten thousand yuan;
(6) water power: annual about 0.8 ten thousand yuan * 5 years=40,000 yuan.
Comprehensively above-mentioned, accumulative 41.2 ten thousand yuan of invention input of expecting funds.
(1) the present invention need choose enter diapause status red shoulder ladybug as diapause maintenance object, the red shoulder ladybug entering diapause status can be obtained by Diapause inducement method, for the determination of red shoulder ladybug Diapause inducement method, applicant has done following experiment.
One, the determination of sensitive period is regulated and controled
Red shoulder ladybug is holometabola predatory natural enemy insect, total ovum, larva, pupa and these four worm states (i.e. developmental stage) of adult in growth course, and the development characteristics of each worm state is far from each other, as ovum, pupa are not eaten motionless; Larva can only take food in aphid pin main body, and mobility is very poor; Adult then has a wing, can circle in the air flexibly.Actually to the red shoulder ladybug of which worm state carry out diapause regulation and control more effectively, easier, need to carry out a series of further investigation.
Through field investigation and in conjunction with lab screening experiment, find that the life stages sensitive that red shoulder ladybug experiences diapause signal is first emerging adult, determine the sensitive period using this stage as Diapause inducement, carry out developmental regulation.
Experiment 1: regulation and control screening test on responsive opportunity
At 15 DEG C, after giving long illumination (14hr/d) and short photoperiod (8hr/d) process by the different developmental phases of red shoulder ladybug respectively by various combination, result shows: the different developmental phases before adult, no matter carry out what kind of short photoperiod process, most emerging adult is energy normal spawning all, diapause does not occur; And adult stage and all developmental stage are when being all placed under short photoperiod condition, most of female worm then can not lay eggs, and enters diapause status.
Table 1 red shoulder ladybug different developmental phases experiences the Diapause rate (15 DEG C) of long illumination (L:14L: 10D) and short photoperiod (S:8L: 16D)
Note: (L:14L: 10D) to refer in 24 hours illumination 14 hours, (S:8L: 16D) to refer in 24 hours illumination 8 hours.
Research shows, the adult eclosion of red shoulder ladybug is the crucial life stages sensitive of its diapause, and under only having adult to be in short photoperiod condition, adult just can enter diapause; Four-age larva, mature larva and the short photoperiod in pupa time experience occurs also there is certain facilitation to the diapause of adult.
Be ovary and the ovary canal of the female worm of diapause red shoulder ladybug as shown in Figure 1 and Figure 2; Fig. 3, the ovary that Figure 4 shows that the female worm of non-diapause and ovary canal.As can be seen from Fig. 1, Fig. 2, ovary and the ovary of the female worm of diapause red shoulder ladybug are still in the non-developmental stage.
Two, the envirment factor combination of Diapause inducement
By regulation and control photoperiod (that is: every day light irradiation time) and temperature, the Diapause inducement of red shoulder ladybug can be realized.Screening test result shows that red shoulder grandis adults is temperature 13 DEG C, photoperiod L8:D16, inducing sustained 192h (that is: always inducing duration) at warm light combination condition, and the individuality of red shoulder ladybug population more than 90% can be induced to enter diapause status.
Experiment 2: Diapause inducement is tested
1. materials and methods:
1.1 for examination worm source and raising method
Red shoulder ladybug picks up from proving ground, Chinese Academy of Agricultural Sciences Langfang wheatland, is formed after the stable generation as supplying examination worm source through indoor feeding.Ovum, pupa and imago breeding condition are temperature 24 ± 1 DEG C, relative moisture 70 ± 10%, photoperiod L16:D8; Larva is fed in dependent insect cage, and condition is temperature 22 DEG C-26 DEG C, relative moisture is 50%-75%, the photoperiod is L:D=16:8.Foodstuff is soybean aphid.
The Diapause inducement effect of 1.2 photoperiods, temperature
If the replica test of each 4 levels of two factors, carry out in climatic cabinate (south of the River, Ningbo RZX500C type), 4 photoperiod levels are L14:D10, L12:D12, L10:D14 and L8:D16 respectively, and 5 temperature levels are 24 DEG C, 21 DEG C, 18 DEG C, 15 DEG C and 13 DEG C respectively; Relative moisture 70 ± 10%.Will the adult male and female of just sprout wings (in 12 hours) match after raise in the plastic cup being equipped with folded sheet (diameter 7cm, high 10cm), rim of a cup cover with gauze in case examination worm escape.Often process repetition 3 times, coprocessing adult 45 is right.Day by day observe and change feed and the scraps of paper, the preoviposition period of recording female worm and individual number of laying eggs, until off-test.
The impact of 1.3 Diapause inducement durations
On the basis of aforementioned 1.1 and 1.2 result of the tests, the temperature determining to be suitable for, photoperiod, carry out in climatic cabinate (south of the River, Ningbo RZX500C type), respectively 48 carried out to red shoulder ladybug, 94,144,192,240, the successive induction of 288h; Relative moisture 70 ± 10%.Will the adult male and female of just sprout wings (in 12 hours) match after raise in the plastic cup being equipped with folded sheet (diameter 7cm, high 10cm), rim of a cup cover with gauze in case examination worm escape.Often process repetition 3 times, coprocessing adult 45 is right.Day by day observe and change feed and the scraps of paper, the preoviposition period of recording female worm and individual number of laying eggs, until off-test.
2. results and analysis:
2.1 photoperiods are on the impact of Diapause inducement
Under different photoperiod condition, the Diapause rate of red shoulder grandis adults is as shown in table 2, under uniform temperature condition, under identical temperature condition, imaginal diapause rate raises gradually with the shortening of light application time generally, under different photoperiod condition adult Diapause rate between there is significant difference.
The table 2 temperature and light cycle is on the impact of red shoulder ladybug Diapause inducement
Note: in table, data are mean value ± standard errors, same column different letter representation Tukey ' s HSD checks significant difference (P<0.05).
When temperature is greater than 18 DEG C, under each illumination, do not occur that diapause is individual; Along with temperature continues to decline, when light irradiation time foreshortens to 8h, occur that diapause is individual; When temperature continues to drop to 15 DEG C, as can be seen from Table 2, the Diapause rate of adult increases gradually with the shortening of light irradiation time, increases to 61.5% of 8h, add 7.7 times from 7.1% of 14h.When temperature drops to 13 DEG C further, under each light irradiation time, the Diapause rate of adult is all higher, total trend still increase with the shortening of light irradiation time, when light irradiation time is 10h, 8h, imaginal diapause rate is respectively 91.7%, 93.8%.Therefore, when carrying out the practical operation of inducing storage to red shoulder ladybug product, the light irradiation time process of 8h-10h can be adopted, red shoulder ladybug can be induced to enter diapause.
2.2 temperature are on the impact of Diapause inducement
Experimental result (table 2) shows, temperature also has remarkable impact to imaginal diapause induction, imaginal diapause rate significantly raises with the decline of temperature (P<0.05), all occurs without diapause individuality under high temperature (21 DEG C and 24 DEG C); Start when being reduced to 18 DEG C with temperature to occur that diapause is individual, but Diapause rate is very low, is only 2.5% (8L: 16D); When temperature is reduced to 15 DEG C, Diapause rate is 61.5%, and photoperiodic Diapause rate more same as 18 DEG C adds 23.6 times; When temperature drops to 13 DEG C further, Diapause rate reaches 93.8%, and most individuality enters diapause status.
Under other photoperiod conditions (14L: 10D, 12L: 12D and 10L: 14D), Diapause rate also reduces with temperature and rises.In addition, temperature also plays obvious regulating action to the critical photoperiod that imaginal diapause is induced.When 15 DEG C, the critical light irradiation time of imaginal diapause induction is between 8 ~ 10hr/d.
To sum up describing low temperature treatment can induce red shoulder ladybug to enter diapause, and high temperature inhibits diapause to occur, and promotes the normal development of ladybug.Under short photoperiod 13 DEG C, 15 DEG C of conditions, the Diapause rate of red shoulder ladybug is 61.5%, 93.8%, and Diapause rate is higher, can adopt on producing.
The interaction in 2.3 temperature and light cycles
13 DEG C and 15 DEG C time, photoperiod and temperature show the two-way analysis of variance that imaginal diapause is induced, photoperiod (F=8.420, df=3, P<0.05) and temperature (F=204.978, df=1, P<0.05) all remarkable impact is had on red shoulder ladybug Diapause rate, and the significant interaction of photoperiod and temperature (F=2.408, df=3, P<0.05).When the photoperiod is constant, imaginal diapause rate reduces with temperature and rises, and the change of temperature lower Diapause rate is more obvious.And only in certain temperature range, just there is remarkable impact the photoperiod on diapause.Show thus, in red shoulder ladybug Diapause inducement effect, temperature is dominant factor, plays a decisive role; Photoperiod is association factor, plays complementary effect.Just because of this mutual synergy of photoperiod and temperature, comprehensive regulation red shoulder ladybug enters diapause.
The impact of 2.4 induction times
On the basis of the first two test, determine 13 DEG C, the induction of L8:D16 combination, carry out the induction experiment of different duration respectively, result shows (as table 3), after successive induction 192h, the Diapause rate of red shoulder ladybug can reach 93.58%, and continue to maintain inductive condition, Diapause rate can increase by a small margin.Consider the factor of Financial cost, the proposed combination in production is temperature 13 DEG C, photoperiod L8:D16, successive induction 192h.
Table 3 induces duration on the impact (13 DEG C, L8:D16) of red shoulder ladybug Diapause inducement
Note: in table, data are mean value ± standard errors, same column different letter representation Tukey ' s HSD checks significant difference (P<0.05).
(2) for red determination of takeing on ladybug diapause maintenance and long storage periods method, applicant has done following experiment, and each step of the method all can be verified from following experiment.
Diapause maintenance is the prerequisite and the basis that extend the natural enemy insect product shelf phase, is also natural enemy insect developmental regulation and the key link of producing combination.Through a large amount of tests, confirm 13 DEG C, the combination condition of 2L:22D is the most favourable, diapause storage period of red shoulder ladybug, some individuals diapause can meet Production requirement completely more than 365d storage period more than 300d.
Three, diapause maintenance test (experiment 3)
1. materials and methods:
1.1 for examination worm source and raising method
1.2 photoperiods, temperature are on the impact of diapause duration
From aforementioned (1) part test result, when 18 DEG C, 21 DEG C, 24 DEG C, the Diapause rate of adult is extremely low is even 0,15 DEG C, 14L: 10D Diapause rate is also lower, therefore this test is only added up that warm light combination condition is 13 DEG C, be induced to the female worm diapause duration of diapause status under 8L: 16D condition.Concrete grammar is as follows: first emerging adult is after photoperiod 8L: 16D induces 192h to enter diapause, post-diapause adult is stored under being placed in the condition of following combination respectively: temperature is set to 15 DEG C, 13 DEG C, 10 DEG C, photoperiod is set to 6L:18D, 4L:20D, 2L:22D and 0L:24D, and raising method is the same.Observe every day and record adult Diapause termination and start time of laying eggs, female worm about 30 is observed in each process.
1.3 data analysis
Diapause duration depends on the interval between diapause entry time and diapause end time, in order to the facility tested, this test using adult just emergence time as diapause entry time, female worm lays eggs the mark terminated as diapause, namely using the female worm of diapause preoviposition period as the diapause duration of adult.
Diapause duration data carry out ANOVA variance analysis after logarithm (LOG10) transforms, and carry out significance test through Duncans multiple comparison graph, statistical analysis software is SAS (V8.0).
Fig. 5 does temperature and photoperiod to affect statistical chart to diapause duration mutually.
2 results and analysis
2.1 photoperiods are on the impact of imaginal diapause duration
As can be seen from Figure 5, the regulation and control of photoperiod to the imaginal diapause duration show obvious quantitative response.In 10 DEG C, 13 DEG C and 15 DEG C of temperature condition, the diapause duration of different photoperiod to the female worm of red shoulder ladybug has certain influence, and the diapause duration of female worm is fallen after rising with the prolongation of light irradiation time.Photoperiod, when being 6L: 18D, diapause duration was shorter; Along with the shortening of light application time, diapause duration extends, and when the photoperiod is 2L: 22D, diapause duration reaches most long status; Comparatively speaking, store under full dark condition, adult declines storage period with instead presenting significance.
2.2 temperature are to the regulating and controlling effect of diapause duration
Result shows, under short photoperiod, temperature is extremely remarkable on the impact of imaginal diapause duration, and favors low temperature is in the diapause development of adult, and diapause duration extends (Fig. 5).When temperature is 15 DEG C, diapause duration is relatively the shortest, and the shortest is 135.3d, and the longest is 160.9d; When dropping to 13 DEG C with temperature, diapause period of storage significant prolongation, average out to 318.4d, the duration, maximum was more than 365.0d; Temperature reduces further, and now diapause individual death rate raises, and instead diapause presents reduction trend storage period, and the shortest is 213.5d, and the longest is 264.7d, although can reach the period of storage of 8 months, individual death rate is high.Comprehensive, when 13 DEG C the diapause intensity of photoperiodic induction than 15 DEG C, 10 DEG C time all eager to excel in whatever one does.
2.3 temperature are to the regulating and controlling effect of diapause duration
Can find out significantly from Fig. 5, the combination in temperature and light cycle significantly affects the length of diapause period of storage, namely affects the length of natural enemy insect product shelf phase.
Comprehensive, under short photoperiod, temperature is extremely remarkable on the impact of imaginal diapause duration, and favors low temperature is in the diapause development of adult, and diapause duration extends (Fig. 5).Best diapause maintenance conditional combination is 13 DEG C, 2L:22D, and now the diapause maintenance time of red shoulder ladybug is the longest, and average shelf life is 318.4d, and the duration, maximum was more than 365.0d, can meet this type of natural enemy insect anniversary formula and expand numerous demand.
(3) for the determination of the present invention's red shoulder ladybug Diapause termination method, applicant has done following experiment, and each step of the present invention all can be verified from following experiment.
Diapause termination is the most critical sport technique segment guaranteeing that natural enemy product release time is consistent with the Field Pests emergence period, the red shoulder ladybug termination of diapause of diapause storage period and diapause maintenance state will be in time, it is made to recover vitality, comprise fecundity, prey ability, circle in the air and expand numerous ability, guarantee that natural enemy product plays the usefulness of Control pests.
Through a large amount of screening experiment, prove following regulation and control parameters combination: temperature 24 DEG C, photoperiod 14L:10D, lasting process 72h, can by the red shoulder ladybug termination of diapause of diapause maintenance state.
Four, Diapause termination test (experiment 4)
1.1 material sources: as aforementioned each test
1.2 temperature are on the impact of red shoulder grandis adults Diapause termination
By the result of previous experiments 2 known 13 DEG C, under 8L: 16D condition, red shoulder ladybug Diapause rate is the highest, more than 90%.Therefore the individuality that this test is not laid eggs under choosing this condition after 40d Diapause inducement is as test worm source.Be provided with 13 DEG C altogether, 8L: 16D, 24 DEG C, 14L: 10D, 10L: 14D and 8L: 16D tetra-process, each process observation 30 is to female worm.Observe every day and change feed and for its fan-shaped scraps of paper of laying eggs, the time preoviposition period of recording female worm.
1.3 data analysis
The Diapause termination of adult is laid eggs as mark using female worm, under different temperatures, photoperiod condition the Diapause termination of adult go through the phase be post-diapause adult transfer to female worm lay eggs between the time interval, namely the combination of female worm temperature light transform after preoviposition period.Before data difference analysis, first data preoviposition period of female worm are carried out logarithm (LOG10) and transform, then utilize the ANOVA in SAS V8 statistical software to carry out variance analysis, significance test of difference adopts Duncan ' s newly multiple differential method.
When Fig. 6 is photoperiod L8:D16, temperature affects statistical chart to red shoulder ladybug Diapause termination; When Fig. 7 is 24 DEG C, the photoperiod affects statistical chart to red shoulder ladybug Diapause termination.
2 results and analysis
2.1 temperature are on the impact of red shoulder ladybug Diapause termination
As can be seen from Figure 6, the photoperiod 8L: 16D time, the significant difference preoviposition period of red shoulder ladybug at 13 DEG C and 24 DEG C, namely the time difference of termination of diapause is remarkable.High temperature promotes the rate of development of post-diapause adult, and the time of termination of diapause is short, and when 24 DEG C, red shoulder ladybug is under short photoperiod, and termination of diapause only needs 4.25d.And low temperature suppresses Adult Development, at 13 DEG C, the diapause that dissolves by itself of red shoulder ladybug needs 290.4d, and the time is nearly the former 70 times.
2.2 photoperiods are on the impact of Diapause termination
As shown in Figure 7, under 24 DEG C of conditions, under the different photoperiod, the time difference of the individual termination of diapause of diapause is remarkable, and the Diapause termination that light irradiation time 8h, 10h, 14h are corresponding is gone through the phase and is respectively 4.25,4.07 and 3.39d.Wherein the shortest have 2d, and the longest have 12d.
Comprehensive two test results, 24 DEG C, 14L:10D, continue process 72h by red for the diapause of more than 98% shoulder ladybug termination of diapause, active period can be entered.
Red shoulder ladybug is utilized to prevent eliminating aphis, by the BIOLOGICAL CONTROL effect of occurring in nature, can continue to control aphid damage, aphid density is maintained under economic injury level, in conjunction with chemical emergent management measure that should be acute, fundamentally can contain the harm of aphid class pest, this is of crucial importance for expanding numerous natural enemy insect meaning the anniversary.
Five, diapause artifact evaluation test (experiment 5)
1. materials and methods
1.1 for examination worm source and raising method: as front
1.2 test methods: be temperature 13 DEG C, photoperiod L8:D16, inducing sustained 192h at warm light combination condition, after making red shoulder ladybug enter diapause state, for examination after storage 150d.To the individuality of diapause state, impose the process of temperature 24 DEG C, photoperiod 14L:10D, lasting process 72h, make its complete termination of diapause.Control group: the adult of the first emergence (in 12 hours) of raising under photoperiod L16:D8, temperature 24 DEG C, relative moisture 70 ± 10% condition in contrast.Process 100 female worms, repeat for 3 times.Total egg laying amount, the egg hatching rate preoviposition period of adding up its longevity of male, female worm life-span, female worm respectively, in the duration of laying eggs, 30 days, evaluate diapause artifact index.
1.3 data analyses: utilize the ANOVA in SAS V8 statistical software to carry out variance analysis, significance test of difference adopts Duncan ' s newly multiple differential method.
2 results and analysis
2.1 body diapause artifact evaluating characteristics
Generally, diapause experience has remarkable impact to the parameter history of life of insect, and for improving the vitality of population, diapause individual need pays certain cost, is reflected on the biological propertyes such as the fecundity of post diapause adult, life-span.
Live through the red shoulder grandis adults of diapause, the life-span of female worm and male worm slightly extends, and slightly extend the preoviposition period of female worm after diapause, the duration of laying eggs significantly increases, and in 30 days, total egg laying amount of female worm significantly raises, and the incubation rate of ovum rises.
Above-mentioned description of test, red its vitality of shoulder ladybug after diapause, biological control potential are obtained for lifting, and diapause experience is favourable to biological and ecological methods to prevent plant disease, pests, and erosion product.
The parameter history of life (24 DEG C) of red shoulder ladybug post diapause adult under the table 4 different photoperiod
Note: imaginal diapause inducing treatment is that emerging adult at the beginning of under 13 DEG C of conditions is through photoperiod L8:D16, successive induction 192h.CK represents the adult process that warm light combines 24 DEG C, raises under L16:D8, and in table, data are mean value ± standard errors, and after data, lowercase alphabet shows significant difference (p<0.05).
Described just in order to the present invention is described above, be construed as the present invention and be not limited to above embodiment, meet the various variants of inventive concept all within protection scope of the present invention.
Claims (6)
1. red shoulder ladybug Diapause termination method, it is characterized in that, the method comprises the following steps:
1) choose enter diapause status red shoulder grandis adults as Diapause termination object;
2), under the red shoulder ladybug chosen in step 1) being placed in Diapause termination environment, by the combination of regulation and control light irradiation time every day, temperature, total light irradiation time three, red shoulder ladybug is made to start to lay eggs thus termination of diapause; Described light irradiation time employing every day photoperiod (8 ~ 14) L:(16 ~ 10) D, temperature is 24 DEG C, continues process 72h.
2. release method as claimed in claim 1, is characterized in that, described step 2) middle light irradiation time employing every day photoperiod 10L:14D.
3. release method as claimed in claim 1, is characterized in that, described step 2) in also to regulate and control the humidity of diapause maintenance environment, relative moisture is 70 ± 10%.
4. release method as claimed in claim 1, it is characterized in that, namely the red shoulder ladybug chosen in described step 1) enters diapause status in the first emerging adult stage, and first emerging adult is the adult within just sprouting wings 12 hours.
5. release method as claimed in claim 1, it is characterized in that, described Diapause termination carries out in climatic cabinate.
6. release method as claimed in claim 1, is characterized in that, after described red shoulder ladybug enters diapause status, through the storage duration of 300 ~ 400d.
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