CN101720724A - Long-term storage method of bee species of artificially diapaused and induced Chouioia cunea - Google Patents
Long-term storage method of bee species of artificially diapaused and induced Chouioia cunea Download PDFInfo
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- CN101720724A CN101720724A CN200910220378A CN200910220378A CN101720724A CN 101720724 A CN101720724 A CN 101720724A CN 200910220378 A CN200910220378 A CN 200910220378A CN 200910220378 A CN200910220378 A CN 200910220378A CN 101720724 A CN101720724 A CN 101720724A
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Abstract
The invention discloses a long-term storage method of bee species of an artificially diapaused and induced Chouioia cunea, which is characterized by comprising the following steps of: a, diapause induction: transferring the Chouioia cunea collected from a wild field to a test tube, inoculating bees by using use white chrysalis, tussah pupa, pine moth pupa or corn borer pupa as a substituted host, and respectively placing the inoculated pupa bodies into an illumination culture box at a temperature of cultivation condition of 22-26 DEG C and the moisture of 70-80 percent; controlling the temperature to be 15-20 DEG C and the daylight illumination time to be 0-12 hours in the illumination culture box after the inoculation is carried out for 36 hours so that the short illumination is carried out during the whole larva period to cause the larva to be in a diapause state; and a mark of the diapause: marking the diapaused mature larva to be taupe; and b, preservation condition: placing the pupa bodies with the diapaused larva into the test tube by using a tampon, and storing the pupa bodies in the culture box or a refrigeration house with constant temperature of 2-10 DEG C and constant moisture in a condition avoiding light. The invention has the advantages that the post-diapause is carried out in the mature larva mode at 3 DEG C and stored for 3-4 months, the emergent Chouioia cunea is not degenerated on egg laying amount, sex rate, activity degree and the like, so as to effectively prolong the storage cycle of the bee species by 4-5 times. The post-diapause development is carried out to comparatively and correctly control the bee emergence time to be favorable for production and field release of artificial bee reproduction.
Description
Technical field
The present invention relates to agriculture and forestry biological control of insect pests field, especially a kind of in the process of artificial breeding Chouioia cunea honeybee kind long-term preservation method.
Background technology
In the artificial breeding process of Chouioia cunea, the preservation of kind of honeybee relies on always constantly to be carried out artificial successive transfer culture after the collection wild bee kind and realizes, it is about 1 that per generation is on average gone through the phase, not only to there be the special messenger to adhere to successive transfer culture throughout the year, and can cause the honeybee kind to be degenerated along with cultivating the increasing of algebraically (generally surpass 3-4 generation), can't continue to expand numerous commodity honeybee production of carrying out, have to gather again new honeybee kind and carry out rejuvenation, the applicant finds that after deliberation Chouioia cunea belongs to the longday type insect, in specific sensitive insect attitude, utilize specific temperature and photoperiod can induce Chouioia cunea to enter the diapause state, diapause can make the long-term stasi of Chouioia cunea, producing termination of diapause again in season, carrying out the honeybee kind expands numerous, can save the cost of throughout the year adhering to successive transfer culture, more can effectively delay the speed that the honeybee kind is degenerated, be the desirable new method of Chouioia cunea kind honeybee long preservation.
Summary of the invention
The purpose of this invention is to provide the Chouioia cunea honeybee kind long-term preservation method that a kind of artificial diapause is induced, utilize the characteristic of Chouioia cunea diapause, realize long preservation honeybee kind and can effectively delay the honeybee kind degenerating by artificial induction and artificial termination of diapause.
The objective of the invention is to realize by the following method:
A. diapause is induced: the Chouioia cunea that the field is adopted back be transferred in vitro with white chrysalis, tussah chrysalis, pine caterpillar or corn borer pupa as an alternative the host connect honeybee, the pupal cell that connects honeybee is placed illumination box respectively, and the condition of breeding is temperature 22-26 ℃, humidity 70%-80%; Connect honeybee after 36 hours with illumination box inner control temperature at 15 ℃-20 ℃, day illumination time, whole larval phase, accepted short illumination, makes it enter the diapause state in 0-12 hour; Enter the sign of diapause: the mature larva body colour of diapause is a taupe.
B. storage condition: the pupal cell that will have the diapause larva seals with tampon and is placed in vitro, and lucifuge stores in 2 ℃ of-10 ℃ of constant temperature and humidity incubators or freezer.
C. diapause is removed: 3 ℃ of low temperature treatment were elevated to 25 ℃-31 ℃ gradually with temperature more than 5 days, humidity 70%-80%, and every day, lighting delay number was 14 hours-24 hours.
Good effect of the present invention:
A. with after the diapause of mature larva form 3 ℃ preserved 3-4 month, degenerating can not appear at aspects such as egg laying amount, property ratio, active degrees in the chalcid fly after the emergence, can effectively prolong honeybee kind retention cycle 4-5 times.
B. grow after the diapause and can control out the honeybee time more accurately, help artificial numerous honeybee production and field and discharge.
Embodiment
1. diapause is induced: the Chouioia cunea that the field is adopted back is transferred in vitro with white chrysalis, tussah chrysalis, pine caterpillar or corn borer pupa host as an alternative connect honeybee, the pupal cell that connects honeybee is placed illumination box respectively, the condition of breeding is 22 ℃-26 ℃ of temperature, humidity 70%-80%, connect honeybee after 36 hours with illumination box inner control temperature at 15 ℃-20 ℃, illumination time day was at 0-12 hour, make it enter the diapause state, the mature larva body colour of diapause is a taupe, and diapause mature larva body colour (taupe) is obviously different with the mature larva color (light yellow) of normal development.
2. storage condition: the pupal cell that will have the diapause larva seals with tampon and is placed in vitro, and lucifuge stores in 2 ℃ of-10 ℃ of constant temperature and humidity incubators or freezer.
3. diapause is removed: 3 ℃ of cryopreservations are surpassed the temperature rise to 25 ℃-30 ℃ gradually in 2 days of 5 days diapause honeybee, and every day, lighting delay number was 14-24 hour, humidity 70%-80%.
4. go out the honeybee fate after diapause is removed: under temperature was respectively 18 ℃, 21 ℃, 24 ℃ and 30 ℃ of conditions, every day, lighting delay number was 14-24 hour, and development duration is about 60.2 days 32.3 days, 22.6 days and 13.6 days respectively after the diapause.
Claims (1)
1. Chouioia cunea honeybee kind long-term preservation method that artificial diapause is induced is characterized in that:
A. diapause is induced: the Chouioia cunea that the field is adopted back be transferred in vitro with white chrysalis, tussah chrysalis, pine caterpillar or corn borer pupa as an alternative the host connect honeybee, the pupal cell that connects honeybee is placed illumination box respectively, and the condition of breeding is temperature 22-26 ℃, humidity 70%-80%; Connect honeybee after 36 hours with illumination box inner control temperature at 15 ℃-20 ℃, day illumination time, whole larval phase, accepted short illumination, makes it enter the diapause state in 0-12 hour; Enter the sign of diapause: the mature larva body colour of diapause is a taupe;
B. storage condition: the pupal cell that will have the diapause larva seals with tampon and is placed in vitro, and lucifuge stores in 2 ℃ of-10 ℃ of constant temperature and humidity incubators or freezer.
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CN2009102203781A CN101720724B (en) | 2009-12-04 | 2009-12-04 | Long-term storage method of bee species of artificially diapaused and induced Chouioia cunea |
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CN2009102203781A CN101720724B (en) | 2009-12-04 | 2009-12-04 | Long-term storage method of bee species of artificially diapaused and induced Chouioia cunea |
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CN101720724A true CN101720724A (en) | 2010-06-09 |
CN101720724B CN101720724B (en) | 2012-02-08 |
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CN105296495A (en) * | 2015-11-26 | 2016-02-03 | 天津师范大学 | cDNA full-length sequence of Chouioia cunea Yang Hsp60 (Heat shock protein) and application |
CN105296497A (en) * | 2015-11-26 | 2016-02-03 | 天津师范大学 | cDNA full-length sequence of Chouioia cunea Yang Hsp83 (Heat shock protein) and application |
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CN105296495A (en) * | 2015-11-26 | 2016-02-03 | 天津师范大学 | cDNA full-length sequence of Chouioia cunea Yang Hsp60 (Heat shock protein) and application |
CN105296497A (en) * | 2015-11-26 | 2016-02-03 | 天津师范大学 | cDNA full-length sequence of Chouioia cunea Yang Hsp83 (Heat shock protein) and application |
CN105296496A (en) * | 2015-11-26 | 2016-02-03 | 天津师范大学 | cDNA full-length sequence of Chouioia cunea Yang Hsp70 (Heat shock protein) and application |
CN105660541A (en) * | 2016-02-15 | 2016-06-15 | 山东省农业科学院植物保护研究所 | Method for inducing reproductive diapause of spotted wing drosophila |
CN105660541B (en) * | 2016-02-15 | 2019-03-01 | 山东省农业科学院植物保护研究所 | A method of induction spotted wing drosophila reproductive diapause |
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CN108849755A (en) * | 2018-06-10 | 2018-11-23 | 山西省农业科学院植物保护研究所 | Oriental fruit months larva store method |
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CN111296365B (en) * | 2020-02-25 | 2021-03-12 | 中国农业科学院茶叶研究所 | Artificial breeding method of Chouioia cunea in tea garden |
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