CN104351140B - Red shoulder ladybug Diapause termination method - Google Patents
Red shoulder ladybug Diapause termination method Download PDFInfo
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- CN104351140B CN104351140B CN201410495765.7A CN201410495765A CN104351140B CN 104351140 B CN104351140 B CN 104351140B CN 201410495765 A CN201410495765 A CN 201410495765A CN 104351140 B CN104351140 B CN 104351140B
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- 230000005058 diapause Effects 0.000 title claims abstract description 213
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- 241000238631 Hexapoda Species 0.000 abstract description 31
- 238000011161 development Methods 0.000 abstract description 15
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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
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- 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 discloses a kind of red shoulder ladybug Diapause termination method, this method comprises the following steps：1）The red shoulder grandis adults chosen into diapause status are used as Diapause termination object；2）By step 1）The red shoulder ladybug of middle selection is placed under Diapause termination environment, by regulate and control daily light irradiation time, temperature, total light irradiation time three combination, red shoulder ladybug is started spawning so as to termination of diapause；The daily light irradiation time uses the photoperiod（8~14）L:（16~10）D, temperature is 24 DEG C, persistently handles 72h.The present invention is by manipulating envirment factor, with reference to necessary treatment measures, realize to excellent predatory natural enemy insect --- the red shoulder ladybug development time started is precisely controlled, and termination of diapause rate can reach more than 98%, and technical guarantee is provided for the anniversary large-scale production of red shoulder ladybug.
Description
Technical field
, can be by by the red shoulder wooden dipper of diapause by this method the present invention relates to a kind of red shoulder ladybug Diapause termination method
Worm adult recovers vitality, including fecundity, prey ability, the numerous ability of expansion of circling in the air, it is ensured that natural enemy product plays control evil
The efficiency of worm.
Background technology
The annual crop yield loss caused by agricultural pest of China is 10-15%, realizes that agricultural pest science is controlled
Reason, for ensureing national food security strategy implementation, promoting agricultural high yield that there are important economical, societal benefits.Aphid is agriculture
Industry produces the first major pest, and annual nearly 5,000,000,000 dollars of the loss caused by aphid in the whole world, is currently mostly to be killed dependent on chemistry
Worm agent preventing and treating aphid class harm.
Preventing and treating agricultural pests during, if simply depend on chemical pesticide, it will bring a series of ecologies, environment,
Negative issue in terms of food security.Due to the long-term use of chemical pesticide, some insects have produced the very strong resistance to the action of a drug, many
The natural enemy for planting insect is largely killed, and causes some Pest rampancies to endanger.In addition, chemical pesticide can polluted-water, air and soil
Earth, and human body is enriched to by food chain, endanger population health.Using biotechnology come prevention and control pest and disease damage, just can effectively it keep away
Exempt from disadvantages mentioned above, thus with wide development prospect.It is exactly one of effective biological control means wherein using natural enemy insect.
Currently a bottleneck problem of natural enemy insect industrialization is both at home and abroad：Natural enemy insect expansion numerous cycle is longer, survival period
Shorter, the product export time is difficult consistent with Injurious insect emergence period, often leads to natural enemy before insect occurs with regard to dead, or insect
Natural enemy product is taken place without to can use.So, the development progress of regulation and control natural enemy insect product has important practical significance.Realize
Regulation and control to natural enemy insect product development progress, can utilize the heritability of insect inherently --- diapause just.
Red shoulder ladybug Leis dimidiata (Fabricius) belong to coleoptera ladybirds, come from Palaearctic region, nowadays alive
Boundary the greater part is distributed, China be distributed mainly on Beijing, Jilin, Liaoning, the Inner Mongol, Hebei, Shandong, Henan, Sichuan,
The ground such as Ningxia, Yunnan and Tibet, can take food a variety of aphids (Jing Ying etc., 2002) on crops and fruit tree.Zong Liang Ping etc.
(1988) research finds that red shoulder ladybug likes best predation cotten aphid Aphis gossypii Glover, but to various wheat aphids, jowar
A variety of aphids such as aphid, soybean aphid Aphis craccivora Koch, acacia aphid Aphis robiniae Macchiati also catch
Food.Ladybug generally eating aphid, but red shoulder ladybug is when aphid lacks, and also takes food tetranychid (Wang Yunhua etc., 1984).It is red
Shoulder ladybug is as one of Dominant Natural Enemies of aphid, and its population quantity is big, and reproduction speed is fast, studies its predation functional responses and is intended to section
Learn and evaluate natural enemy control action, scientific basis is provided to better profit from nature enemy in amount.
In recent years, red shoulder ladybug and achieves significant ecological benefits and warp in many countries by successive development and application
Ji benefit, has played irreplaceable prevention and control effect in the biological control of aphid.But in production application, it is commonly present expansion
Numerous cycle is short, product can not for a long time be stored and can not applied in good time, imaginal diapause and the problems such as influence to expand numerous efficiency, as restriction
The technical bottleneck of red shoulder ladybug large-scale production application.
The content of the invention
For problems of the prior art, the invention provides a kind of red shoulder ladybug Diapause termination method, pass through behaviour
Envirment factor is controlled, such as daily light irradiation time, temperature, with reference to necessary treatment measures, are realized to excellent predator elder brother
Worm --- red shoulder ladybug develops being precisely controlled for time started.
The purpose of the present invention is achieved through the following technical solutions：
Red shoulder ladybug Diapause termination method, this method comprises the following steps：
1) the red shoulder grandis adults chosen into diapause status are used as Diapause termination object；
2) by step 1) in choose red shoulder ladybug be placed under Diapause termination environment, by regulating and controlling daily light irradiation time, temperature
Degree, the combination of total light irradiation time three, make red shoulder ladybug start spawning so as to termination of diapause；The daily light irradiation time uses light
Cycle (8~14) L:(16~10) D, temperature is 24 DEG C, persistently handles 72h.Here 72h refers to lasting processing 3 days.
Further, the step 2) in daily light irradiation time use photoperiod 10L:14D.
Further, the step 2) in also the humidity of diapause maintenance environment is regulated and controled, relative humidity be 70 ±
10%.
Further, the step 1) in the red shoulder ladybug chosen in the first emerging adult stage be to enter diapause status, first plumage
Worm is melted into for the adult within just sprouting wings 12 hours.
Further, the Diapause termination is carried out in growth cabinet.
Further, the red shoulder ladybug enters after diapause status, by 300~400d storage duration.
The present invention is using the red shoulder ladybug into diapause status as Diapause termination object, to the time spy useless of diapause maintenance
It is different to require, if using the red shoulder grandis adults of diapause stored for a long time as object is released, following red shoulder ladybug can be used
Diapause maintenance and long-term storage method.
Red shoulder ladybug diapause maintenance and long-term storage method, this method comprise the following steps：
1) the red shoulder ladybug for entering diapause status is chosen as diapause maintenance object, and the red shoulder ladybug of the state sprouts wings just
Adult stage is to enter diapause status；
2) by step 1) in choose red shoulder ladybug be placed under diapause maintenance environment, by regulating and controlling daily light irradiation time, temperature
Both combinations of degree, make red shoulder ladybug maintain diapause status；The daily light irradiation time uses photoperiod (2~6) L:(22~
18) D, temperature is 10~15 DEG C, a length of 300~400d during storage.
Further, the step 2) in daily light irradiation time use photoperiod 2L:22D, temperature is 13 DEG C, storage duration >=
365d。
Further, the step 2) in also the humidity of diapause maintenance environment is regulated and controled, relative humidity be 70 ±
10%.
Further, the step 1) in just emerging adult for just sprout wings 12 hours within adult.
Further, the diapause maintenance and long-term be stored in growth cabinet are carried out.
Red shoulder ladybug into diapause status can be obtained by following Diapause inducement method.
Red shoulder ladybug Diapause inducement method, this method comprises the following steps：
1) sensitive period for determining red shoulder ladybug diapause regulation and control is first emerging adult, chooses the red shoulder ladybug conduct of the worm state
Diapause inducement object；
2) by step 1) in choose red shoulder ladybug be placed under Diapause inducement environment, by regulating and controlling daily light irradiation time, temperature
Degree, the combination of total induction duration three, make red shoulder ladybug enter diapause status；The daily light irradiation time using the photoperiod (8~
10)L:(16~14) D, temperature is 13~15 DEG C, successive induction time >=192h.Here 192h refers to 8 days.
Further, the step 2) in daily light irradiation time be 8hr/d, temperature is 13 DEG C, a length of 192h during total induction.
Further, the step 2) in also the humidity of Diapause inducement environment is regulated and controled, relative humidity be 70 ±
10%.
Further, the step 2) in the successive induction time be 240h or 288h.
Further, the step 1) in just emerging adult for just sprout wings 12 hours within adult.
Further, the Diapause inducement is carried out in growth cabinet.
Further, it is real to four-age larva, mature larva and the pupa time of red shoulder ladybug before Diapause inducement object is chosen
Short photoperiod processing is applied, daily light irradiation time is 8~10hr/d.
There is advantages below using the red shoulder ladybug Diapause termination method of above-mentioned technical characteristic：
The present invention is directed to red this excellent predatory natural enemy insect of shoulder ladybug Leis dimidiata (Fabricius),
The invention research using the cytokine regulatory such as photoperiod, thermoperiod worm development progress is carried out, for Diapause termination system
System research, realizes the control accurate to red shoulder ladybug development progress.The red shoulder ladybug stored with diapause status, its product
30 days of shelf life never diapause are extended to 300 days, the red shoulder ladybug vitality after Diapause termination for example fecundity, egg laying amount,
There are different degrees of raising in spawning duration, life-span etc..Using this technology, the red shoulder ladybug of whole year production can be achieved, will
Its product is stored by diapause technology, is provided natural enemy product for the biological control of aphid class pest, is enriched biological control
Means, improve the science improvement level of agricultural pests.
Carry out the diapause control technique research of red shoulder ladybug, the extensive of red shoulder ladybug can not only be promoted in production practices
Expand it is numerous, extend its shelf life and the prevention and control action time in field, it helps grasp natural enemy insect development characteristicses with
Generation dynamic, improves control of insect efficiency, additionally aids the understanding deepened to natural enemy insect developmental mechanism, explores insect to environment
Adaptation mechanism and evolutionary path.
Brief description of the drawings
Fig. 1 is the ovary displaing micro picture of the red shoulder ladybug female adult of diapause；
Fig. 2 is the ovary canal displaing micro picture of the red shoulder ladybug female adult of diapause；
Fig. 3 is the ovary displaing micro picture of the red shoulder ladybug female adult of non-diapause；
Fig. 4 is the ovary canal displaing micro picture of the red shoulder ladybug female adult of non-diapause；
Fig. 5 is temperature and the photoperiod mutual influence statistical chart made to diapause duration；
Fig. 6 is photoperiod L8：Influence statistical chart of the temperature to red shoulder ladybug Diapause termination during D16；
Influence statistical chart of the photoperiod to red shoulder ladybug Diapause termination when Fig. 7 is 24 DEG C.
Embodiment
Further to illustrate the present invention to reach technological means and effect that predetermined goal of the invention is taken, below in conjunction with
Accompanying drawing and preferred embodiment, structure, workflow to the present invention describe in detail as follows.
The present invention be applicant on the basis of many experiments, expend 5 years cultivated, checking is acquired technology into
Really.
1st, time cost：From 2010 so far, 5 years are lasted.
2nd, financial cost：
(1) equipment：Successively purchase growth cabinet 7 (Ningbo south of the River 500C types), expenditure funds 1.5*7=11.2 ten thousand
Member；
(2) reagent：Purchase the necessary seed of holding test worm population, liquid fertilizer, vermiculite, turf, progress Diapause inducement survey
The necessary chemical reagent of examination, adds up * 5 years=7.5 ten thousand yuan of 1.5 ten thousand yuan/year of funds；
(3) it is artificial：In addition to inventor works, the people of casual labour 2,1200 yuan/month * 10 months * 5 years=6 are still employed
Wan Yuan；
(4) place：Except the experiment using inventor's unit is outdoor, still rents and test greenhouse, solarium, 10,000 yuan of annual cost,
Accumulative 50,000 yuan of funds；
(5) consumptive material：Support the experiment such as worm frame, insect cage, temperature-humidity monitoring instrument, photoperiod monitor, time switch, sunshade net
Consumptive material, adds up * 5 years=7.5 ten thousand yuan of 1.5 ten thousand yuan/year；
(6) water power：It is annual about 0.8 ten thousand yuan * 5 years=40,000 yuan.
Summary, invention input of expecting funds are accumulative 41.2 ten thousand yuan.
(1) present invention needs to choose the red shoulder ladybug for entering diapause status as diapause maintenance object, into diapause status
Red shoulder ladybug can be obtained by Diapause inducement method, for the determination of red shoulder ladybug Diapause inducement method, applicant does
Test below.
First, the determination of sensitive period is regulated and controled
Red shoulder ladybug be have in holometabola predatory natural enemy insect, growth course ovum, larva, pupa and adult this four
Individual worm state (i.e. stage of development), the development characteristicses of each worm state are far from each other, and such as ovum, pupa does not eat motionless；Larva can only be in aphid
Taken food in pin main body, mobility is very poor；Adult then has wing, can flexibly circle in the air.Actually to the red shoulder ladybug of which worm state
Carry out diapause regulation and control more effective, easier, it is necessary to carry out a series of further investigation.
Through field investigation and combination lab screening experiment, it is found that the life stages sensitive that red shoulder ladybug experiences diapause signal is first plumage
It is melted into worm, it is determined that using this stage as the sensitive period of Diapause inducement, carries out developmental regulation.
Experiment 1：The sensitive opportunity screening test of regulation and control
At 15 DEG C, long illumination (14hr/d) is given respectively by various combination and short by the different developmental phases of red shoulder ladybug
After illumination (8hr/d) processing, as a result show：What kind of short photoperiod processing no matter different developmental phases before adult, carry out,
Most emerging adults energy normal spawning, does not occur diapause；And adult stage and all stages of development are placed in short photoperiod bar
When under part, most of female adult can not then lay eggs, into diapause status.
The red shoulder ladybug different developmental phases of table 1 experience long illumination (L：And short photoperiod (S 14L: 10D)：Diapause 8L: 16D)
Rate (15 DEG C)
Note：(L：Refer to 24 hour in illumination 14 hour, (S 14L: 10D)：Refer to 24 hour in illumination 8 hour 8L: 16D).
Research shows that the adult eclosion of red shoulder ladybug is the crucial life stages sensitive of its diapause, and only adult be in it is short
Under illumination condition, adult could enter diapause；The diapause of four-age larva, mature larva and the short photoperiod in pupa time experience to adult is sent out
Life also has certain facilitation.
It is the ovary and ovary canal of the red shoulder ladybug female adult of diapause as shown in Figure 1 and Figure 2；It is female that Fig. 3, Fig. 4 show non-diapause
The ovary and ovary canal of worm.It is still within not sending out from Fig. 1, Fig. 2 ovary and ovary that can be seen that the red shoulder ladybug female adult of diapause
Educate the stage.
2nd, the envirment factor combination of Diapause inducement
By regulating and controlling the photoperiod (i.e.：Daily light irradiation time) and temperature, the Diapause inducement of red shoulder ladybug can be achieved.Screening examination
Test result and show that red shoulder grandis adults are 13 DEG C of temperature, photoperiod L8 in warm light combination condition:D16, inducing sustained 192h are (i.e.：
Total induction duration), it can induce red individual of the shoulder ladybug population more than 90% and enter diapause status.
Experiment 2：Diapause inducement is tested
1. materials and methods：
1.1 selected insect sources and method for breeding
Red shoulder ladybug picks up from Chinese Academy of Agricultural Sciences Langfang proving ground wheatland, is after forming the stable generation through indoor feeding
Selected insect source.Ovum, pupa and imago breeding condition are 24 ± 1 DEG C of temperature, relative humidity 70 ± 10%, photoperiod L16:D8；Larva
In being fed in insect cage, condition be 22 DEG C -26 DEG C of temperature, relative humidity be 50%-75%, photoperiod be L:D=16:8.Foodstuff
For soybean aphid.
1.2 photoperiods, the Diapause inducement effect of temperature
If the replica test of each 4 level of two factors, in progress, 4 in growth cabinet (Ningbo south of the River RZX500C types)
Photoperiod level is L14 respectively:D10、L12:D12、L10:D14 and L8:D16,5 temperature levels are 24 DEG C respectively, 21 DEG C, 18
DEG C, 15 DEG C and 13 DEG C；Relative humidity 70 ± 10%.By just sprout wings (in 12 hours) the pairing of adult male and female after raise in being equipped with folding
In the plastic cup (diameter 7cm, high 10cm) of the scraps of paper, rim of a cup cover is escaped with gauze to prevent test worm.Often processing is repeated 3 times, coprocessing
45 pairs of adult.Day by day observe and change feed and the scraps of paper, record female adult preoviposition period and spawning number of individuals, until off-test.
The influence of 1.3 Diapause inducement durations
On the basis of foregoing 1.1 and 1.2 result of the tests, it is determined that suitable temperature, photoperiod, (peaceful in growth cabinet
Ripple south of the River RZX500C types) in carry out, carry out 48 to red shoulder ladybug respectively, 94,144,192,240,288h successive induction；Phase
To humidity 70 ± 10%.By just sprout wings (in 12 hours) the pairing of adult male and female after raise it is (straight in the plastic cup for being equipped with folded sheet
Footpath 7cm, high 10cm) in, rim of a cup cover is escaped with gauze to prevent test worm.Often processing is repeated 3 times, 45 pairs of coprocessing adult.Day by day see
Examine and change feed and the scraps of paper, record female adult preoviposition period and spawning number of individuals, until off-test.
2. result and analysis：
Influence of 2.1 photoperiods to Diapause inducement
The Diapause rate of red shoulder grandis adults is as shown in table 2 under the conditions of the different photoperiods, under the conditions of certain temperature, phase equality of temperature
Under the conditions of degree, imaginal diapause rate generally gradually rises with the shortening of light application time, and adult is stagnant under the conditions of the different photoperiods
Educate between rate and there is significant difference.
The influence of the temperature of table 2 and photoperiod to red shoulder ladybug Diapause inducement
Note：Data are average value ± standard errors in table, and the different letters of same column represent that Tukey ' s HSD examine significant difference (P
<0.05)。
When temperature is more than 18 DEG C, do not occur diapause individual under each illumination；As temperature continues to decline, light irradiation time contracting
It is as short as occurring diapause individual during 8h；When temperature continues to drop to 15 DEG C, from table 2 it can be seen that the Diapause rate of adult is with illumination
The shortening of duration and gradually increase, increase to the 61.5% of 8h from the 7.1% of 14h, add 7.7 times.When temperature further declines
During to 13 DEG C, the Diapause rate of adult is all higher under each light irradiation time, and total trend still increases with the shortening of light irradiation time,
When light irradiation time is 10h, 8h, imaginal diapause rate is respectively 91.7%, 93.8%.Therefore lured to red shoulder ladybug product
When leading the practical operation of storage, it can be handled using 8h-10h light irradiation time, can induce red shoulder ladybug and enter diapause.
Influence of 2.2 temperature to Diapause inducement
Experimental result (table 2) shows that temperature is induced imaginal diapause also to have and significantly affected, and imaginal diapause rate is with temperature
Decline and significantly raise (P<0.05), occur under high temperature (21 DEG C and 24 DEG C) without diapause individual；When being reduced to 18 DEG C with temperature
Start diapause individual occur, but Diapause rate is very low, only 2.5% (8L: 16D)；When temperature is reduced to 15 DEG C, Diapause rate is
61.5%, the Diapause rate of photoperiod more same than 18 DEG C adds 23.6 times；When temperature further drops to 13 DEG C, Diapause rate reaches
93.8%, most individuals enter diapause status.
Under other photoperiod conditions (14L: 10D, 12L: 12D and 10L: 14D), Diapause rate is also reduced and risen with temperature.
In addition, the critical photoperiod that temperature is induced imaginal diapause also functions to obvious adjustment effect.Imaginal diapause is induced at 15 DEG C
Critical light irradiation time is between 8~10hr/d.
To sum up illustrate that low-temperature treatment can induce red shoulder ladybug and enter diapause, and high temperature inhibits diapause, promotes wooden dipper
The normal development of worm.Under the conditions of 13 DEG C, 15 DEG C of short photoperiod, the Diapause rate of red shoulder ladybug is 61.5%, 93.8%, Diapause rate compared with
It can be used in height, production.
2.3 temperature and the reciprocation of photoperiod
At 13 DEG C and 15 DEG C, the two-way analysis of variance that photoperiod and temperature are induced imaginal diapause shows, photoperiod (F
=8.420, df=3, P<0.05) with temperature (F=204.978, df=1, P<0.05) have significantly to red shoulder ladybug Diapause rate
Influence, and significant interaction (F=2.408, df=3, the P of photoperiod and temperature<0.05).When the photoperiod is constant, into
Worm Diapause rate is reduced and risen with temperature, and temperature is lower, and Diapause rate change is more obvious.And only in certain temperature range, light week
Phase just has a significant impact to diapause.It is indicated above that in red shoulder ladybug Diapause inducement effect, temperature is dominant factor, rises and determine
Property effect；Photoperiod is association factor, is served complementary.Just because of photoperiod and this mutual synergy of temperature,
The red shoulder ladybug of comprehensive regulation enters diapause.
The influence of 2.4 induction times
On the basis of the first two experiment, it is determined that 13 DEG C, L8:D16 induction combination, carries out luring for different durations respectively
Experiment is led, is as a result shown after (such as table 3), successive induction 192h, the Diapause rate of red shoulder ladybug continues to lure up to 93.58%
Sliver part, Diapause rate can increase by a small margin.In view of the factor of financial cost, the proposed combination in production is 13 DEG C of temperature, light
Period L 8:D16, successive induction 192h.
Table 3 induces influence (13 DEG C, L8 of the duration to red shoulder ladybug Diapause inducement:D16)
Note：Data are average value ± standard errors in table, and the different letters of same column represent that Tukey ' s HSD examine significant difference (P
<0.05)。
(2) for red shoulder ladybug diapause maintenance and the determination of long-term storage method, applicant has done following experiment, this method
Each step can be verified from following experiment.
Diapause maintenance is premise and the basis for extending natural enemy insect shelf life, is also natural enemy insect developmental regulation and life
Produce the key link combined.By substantial amounts of experiment, it was demonstrated that 13 DEG C, 2L:22D combination condition is the most favourable, red shoulder ladybug
Diapause storage period, more than 365d, can meet production requirement completely more than 300d, some individuals diapause storage period.
3rd, diapause maintenance experiment (experiment 3)
1. materials and methods：
1.1 selected insect sources and method for breeding
The influence of 1.2 photoperiods, temperature to diapause duration
From foregoing (1) part test result, at 18 DEG C, 21 DEG C, 24 DEG C, the Diapause rate extremely low even 0 of adult,
15 DEG C, 14L: 10D Diapause rate it is relatively low, therefore this experiment only count warm light combination condition be 13 DEG C, induce under the conditions of 8L: 16D to
The female adult diapause duration of diapause status.Specific method is as follows：First emerging adult enters through the induction of photoperiod 8L: 16D 192h
After diapause, post-diapause adult is stored under conditions of being respectively placed in following combination：Temperature setting is 15 DEG C, 13 DEG C, 10 DEG C, and the photoperiod sets
It is set to 6L:18D、4L:20D、2L:22D and 0L:24D, method for breeding is the same.Observe daily and record adult Diapause termination and open
Begin the time laid eggs, each processing observation female adult 30 or so.
1.3 data analysis
Diapause duration depends on the interval between diapause entry time and diapause end time, for experiment just
Profit, the mark that this experiment terminates emergence time at the beginning of adult as diapause entry time, female adult spawning as diapause, i.e., with diapause
The preoviposition period of female adult as adult diapause duration.
Diapause duration data carry out ANOVA variance analyses after being converted through logarithm (LOG10), through Duncans multiple ratios
Significance test is carried out compared with method, statistical analysis software is SAS (V8.0).
Fig. 5 is temperature and the photoperiod mutual influence statistical chart made to diapause duration.
2 results and analysis
Influence of 2.1 photoperiods to the imaginal diapause duration
From fig. 5, it can be seen that regulation and control of the photoperiod to the imaginal diapause duration show obvious quantitative response.10
DEG C, in 13 DEG C and 15 DEG C of temperature conditionss, there are certain influence the different photoperiods to the diapause duration of red shoulder ladybug female adult, female
The diapause duration of worm is fallen after rising with the extension of light irradiation time.When photoperiod is 6L: 18D, diapause duration is shorter；With
The shortening of light application time, diapause duration extends, when the photoperiod is 2L: 22D, diapause duration reaches most long shape
State；Comparatively, stored under completely black dark condition, adult storage period declines while conspicuousness is instead presented.
Regulating and controlling effect of 2.2 temperature to diapause duration
As a result show, under short photoperiod, temperature influences extremely notable to the imaginal diapause duration, and favors low temperature is in adult
Diapause development, diapause duration extension (Fig. 5).When temperature is 15 DEG C, diapause duration is relatively most short, most short to be
135.3d, up to 160.9d；During with temperature drop to 13 DEG C, diapause period of storage significantly extends, average out to 318.4d, continues
Time, maximum was more than 365.0d；Temperature is further reduced, and instead now diapause individual death rate rise, diapause storage period is presented
Reduction trend, most short is 213.5d, up to 264.7d, although the period of storage of 8 months can be reached, individual death rate
It is high.In terms of comprehensive, 13 DEG C when photoperiodic induction diapause intensity than 15 DEG C, 10 DEG C when will be strong.
Regulating and controlling effect of 2.3 temperature to diapause duration
It can be apparent from from Fig. 5, the combination of temperature and photoperiod significantly affect the length of diapause period of storage, i.e. shadow
Ring the length of natural enemy insect shelf life.
In terms of comprehensive, under short photoperiod, temperature influences extremely notable to the imaginal diapause duration, and favors low temperature is in adult
Diapause development, diapause duration extension (Fig. 5).Optimal diapause maintenance conditional combination is 13 DEG C, 2L:22D, now red shoulder wooden dipper
The diapause maintenance time of worm is most long, and average shelf life is 318.4d, and duration maximum can meet such day more than 365.0d
Oppose insect anniversary formula and expand numerous demand.
(3) determination of red shoulder ladybug Diapause termination method of the invention is directed to, applicant has done following experiment, and of the invention is each
Individual step can be verified from following experiment.
Diapause termination is to ensure that the natural enemy product release time most critical sport technique segment consistent with the Field Pests generation phase, and
When by be in diapause storage period diapause maintenance state red shoulder ladybug termination of diapause, its is recovered vitality, including reproduction
Ability, prey ability, the numerous ability of expansion of circling in the air, it is ensured that natural enemy product plays the efficiency of control insect.
By a large amount of screening experiments, it was demonstrated that following regulation and control parameter combination：24 DEG C of temperature, photoperiod 14L:10D, lasting place
Manage 72h, you can by the red shoulder ladybug termination of diapause of diapause maintenance state.
4th, Diapause termination experiment (experiment 4)
1.1 material source：Such as foregoing each experiment
Influence of 1.2 temperature to red shoulder grandis adults Diapause termination
13 DEG C, under the conditions of 8L: 16D, red shoulder ladybug Diapause rate highest, more than 90% are understood by the result of previous experiments 2.Therefore
This experiment chooses the individual do not laid eggs after 40d Diapause inducements under the conditions of this as experiment worm sources.Altogether provided with 13 DEG C, 8L: 16D,
24 DEG C, 14L: 10D, 10L: 14D and 8L: 16D tetra- processing, 30 pairs of female adults of each processing observation.Daily observe and change feed and
For the fan-shaped scraps of paper of its spawning, female adult time preoviposition period is recorded.
1.3 data analysis
The Diapause termination of adult using female adult spawning as indicate, different temperatures, under the conditions of the photoperiod adult Diapause termination
It is the time interval that post-diapause adult is transferred between female adult spawning to go through the phase, i.e. preoviposition period after female adult temperature light combination conversion.Number
Before variance analysis, female adult preoviposition period data are first subjected to logarithm (LOG10) and converted, then using in SAS V8 statistical softwares
ANOVA carry out variance analysis, significance test of difference uses Duncan ' s newly multiple differential methods.
Fig. 6 is photoperiod L8：Influence statistical chart of the temperature to red shoulder ladybug Diapause termination during D16；Fig. 7 is 24 DEG C of weeks in time
Influence statistical chart of the phase to red shoulder ladybug Diapause termination.
2 results and analysis
Influence of 2.1 temperature to red shoulder ladybug Diapause termination
As can be seen from Figure 6, in photoperiod 8L: 16D, red shoulder ladybug preoviposition period significant difference, that is, solve at 13 DEG C and 24 DEG C
Except the time difference of diapause is notable.High temperature promotes the rate of development of post-diapause adult, and the time of termination of diapause is short, at 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, red shoulder ladybug solves naturally
Except diapause needs 290.4d, the time is nearly the former 70 times.
Influence of 2.2 photoperiods to Diapause termination
As shown in Figure 7, under the conditions of 24 DEG C, the time difference of diapause individual termination of diapause is not notable under the different photoperiods,
It is respectively 4.25,4.07 and 3.39d that the corresponding Diapause termination of light irradiation time 8h, 10h, 14h, which goes through the phase,.It is wherein most short to have a 2d, it is most long
There is 12d.
The result of comprehensive two experiments, 24 DEG C, 14L:10D, persistently processing 72h can be by the red shoulder ladybug of more than 98% diapause
Termination of diapause, into active period.
Eliminate aphis, acted on by the BIOLOGICAL CONTROL in nature using red shoulder ladybug is anti-, control aphid damage can be continued, will
Aphid density is maintained under economic injury level, with reference to acute chemical emergent management measure is answered, can fundamentally be contained
The harm of aphid class pest, it is of crucial importance that this expands numerous natural enemy insect meaning for the anniversary.
5th, biological assessment experiment (experiment 5) after diapause
1. materials and methods
1.1 selected insect sources and method for breeding：As before
1.2 test method：It is 13 DEG C of temperature, photoperiod L8 in warm light combination condition:D16, inducing sustained 192h, make red shoulder
Ladybug enters after diapause state, for examination after storage 150d.To the individual of diapause state, 24 DEG C of temperature, photoperiod 14L are imposed:10D, hold
Continuous processing 72h processing, makes its complete termination of diapause.Control group：Photoperiod L16:D8,24 DEG C of temperature, relative humidity 70 ±
The adult for the first emergence (in 12 hours) raised under the conditions of 10% is as control.Handle 100 female adults, 3 repetitions.Count respectively
Its longevity of male, female adult life-span, female adult preoviposition period, total egg laying amount in the spawning duration, 30 days, egg hatching rate, are evaluated stagnant
Educate rear biological indicator.
1.3 data analysis：Variance analysis is carried out using the ANOVA in SAS V8 statistical softwares, significance test of difference is adopted
Differential method is newly answered with Duncan ' s.
2 results and analysis
Biological evaluation after 2.1 body diapause
Generally, diapause experience has on the history of life parameter of insect and significantly affected, to improve the vitality of population,
Diapause individual need pays certain cost, is reflected on the biological characteristicses such as the fecundity of post diapause adult, life-span.
The red shoulder grandis adults of diapause are lived through, the life-span of female adult and male worm slightly has extension, before the female adult spawning after diapause
Phase slightly has extension, and the spawning duration is dramatically increased, and total egg laying amount of female adult is significantly raised in 30 days, and the incubation rate of ovum rises.
Above-mentioned description of test, red its vitality of shoulder ladybug, biological control potential after diapause are obtained for lifting,
Diapause experience is favourable to biological and ecological methods to prevent plant disease, pests, and erosion product.
The history of life parameter (24 DEG C) of red shoulder ladybug post diapause adult under the different photoperiods of table 4
Note：Imaginal diapause inducing treatment is first emerging adult under the conditions of 13 DEG C through photoperiod L8:D16, successive induction
192h.CK represents that warm light combines 24 DEG C, L16:Data are average value ± standard error, data in the adult processing raised under D8, table
Afterwards lowercase letter indication difference significantly (p<0.05).
It is described above simply to illustrate that of the invention, it is understood that the invention is not limited in above example, meet
The various variants of inventive concept are within protection scope of the present invention.
Claims (4)
1. red shoulder ladybug Diapause termination method, it is characterised in that this method comprises the following steps：
1) the red shoulder grandis adults chosen into diapause status are used as Diapause termination object；
2) by step 1) in the red shoulder ladybug chosen be placed under Diapause termination environment, by regulating and controlling daily light irradiation time, temperature, total
The combination of light irradiation time three, makes red shoulder ladybug start spawning so as to termination of diapause；The daily light irradiation time uses the photoperiod
10L:14D, temperature is 24 DEG C, persistently handles 72h；
The step 2) in also the humidity of diapause maintenance environment is regulated and controled, relative humidity be 70 ± 10%.
2. release method as claimed in claim 1, it is characterised in that the step 1) in the red shoulder ladybug chosen sprout wings just
Adult stage is to enter diapause status, and first emerging adult is the adult within just sprouting wings 12 hours.
3. release method as claimed in claim 1, it is characterised in that the Diapause termination is carried out in growth cabinet.
4. release method as claimed in claim 1, it is characterised in that the red shoulder ladybug enters after diapause status, by 300
~400d storage duration.
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