CN106376528B - A kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao - Google Patents
A kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao Download PDFInfo
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- 206010039710 Scleroderma Diseases 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 18
- 241000382353 Pupa Species 0.000 claims abstract description 33
- 239000000463 material Substances 0.000 claims abstract description 11
- 238000011161 development Methods 0.000 claims abstract description 10
- 230000018109 developmental process Effects 0.000 claims abstract description 10
- 238000012545 processing Methods 0.000 claims abstract description 7
- 230000036541 health Effects 0.000 claims abstract description 5
- 230000032669 eclosion Effects 0.000 claims abstract description 4
- 241000238631 Hexapoda Species 0.000 abstract description 2
- 241001481710 Cerambycidae Species 0.000 description 9
- 102000002322 Egg Proteins Human genes 0.000 description 9
- 108010000912 Egg Proteins Proteins 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 210000004681 ovum Anatomy 0.000 description 9
- 241001442207 Monochamus alternatus Species 0.000 description 7
- 238000009395 breeding Methods 0.000 description 6
- 230000001488 breeding effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000008014 freezing Effects 0.000 description 5
- 238000007710 freezing Methods 0.000 description 5
- 238000004781 supercooling Methods 0.000 description 5
- 241000243771 Bursaphelenchus xylophilus Species 0.000 description 4
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 4
- 235000011613 Pinus brutia Nutrition 0.000 description 4
- 241000018646 Pinus brutia Species 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 4
- -1 small molecule alcohols Chemical class 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- 229920002527 Glycogen Polymers 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- 241000244206 Nematoda Species 0.000 description 3
- 235000019784 crude fat Nutrition 0.000 description 3
- 229940096919 glycogen Drugs 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 230000003071 parasitic effect Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 244000050510 Cunninghamia lanceolata Species 0.000 description 2
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- 230000022472 cold acclimation Effects 0.000 description 2
- 244000078703 ectoparasite Species 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241001042196 Batocera horsfieldi Species 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- 241001116468 Cunninghamia Species 0.000 description 1
- 241000254171 Curculionidae Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000282376 Panthera tigris Species 0.000 description 1
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- 241001079064 Zanthoxylum schinifolium Species 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
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- 238000004891 communication Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003933 environmental pollution control Methods 0.000 description 1
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- 239000000835 fiber Substances 0.000 description 1
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- 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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Abstract
The present invention relates to insect domestication by low temperature method fields, specially a kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao.The method comprising the steps of 1), domestication kind bee selects: selecting room temperature to cultivate, the advanced age pupa (10 ~ 14d after pupating) of development health is as domestication material；2), domestication by low temperature is handled: setting 7 ~ 9h of domestication at 8 DEG C of 10 ~ ﹣ of ﹣；3) culture under room temperature, orientation domestication: is put back to after first generation advanced age pupa domestication by low temperature at bee eclosion, it takes into survive female and carries out next-generation bee at room temperature at bee and cultivate, it is developed to the pupa stage at advanced age to bee and carries out domestication by low temperature processing, continuous domestication n(n≤8 again) generation.The Scleroderma sichuanensis Xiao cold resistance that this method obtains significantly increases, and according to said method orientation, which is tamed, can get the stronger Scleroderma sichuanensis Xiao low temperature strain of cold resistance.
Description
Technical field
The present invention relates to insect domestication by low temperature method fields, specially a kind of domestication by low temperature side in short-term of Scleroderma sichuanensis Xiao
Method.
Background technique
Scleroderma sichuanensis Xiao Scleroderma sichuanensis Xiao is the double China firs of the thick sheath of parasitism of discovery in 1994
The ectoparasite bee of roundheaded borer and pupa.Host range is wide for the bee, attack is strong for search, parasitic rate is high, is that a kind of can effectively prevent
The excellent parasitic wasp of the forests borer pest such as part longicorn, bark beetle, buprestid beetle and weevil.Wherein with the control effect to longicorn
It is most significant.The discovery of the bee and extensive artificial breeding application are " research of China fir longicorn integrated control technique " and " economic forest brill moth
The master of two scientific achievements of pest non-environmental pollution control technical research " (Sichuan provincial government, scientific and technological progress second prize in 1998,2005)
Component part is wanted, has made a breakthrough, can have been produced in batches throughout the year in the extensive artificial breeding technology field of swollen leg bee.
Studies have shown that Scleroderma sichuanensis Xiao is not only control China fir palm fibre longicorn, Semanotus bifasciatus sinoauster, Chinese prickly ash tiger longicorn, blueness
The ideal natural enemy of the middle-size and small-size longicorn such as Yang Tianniu and apple eye longicorn, and it is medium-and-large-sized to Monochamus alternatus Hope, longicorn beetle, batocera horsfieldi etc.
The control effect of longicorn is also more significant.Pine nematode is a kind of destructiveness forest disease, contagiousness as caused by Bursaphelenchus xylophilus
By force, lethal speed is fast, difficulty of prevention and cure is big, is known as " cancer " of pine tree.Monochamus alternatus Hope is the key that Bursaphelenchus xylophilus communication media,
It reduces and control Monochamus alternatus Hope population quantity, effective cutting Bursaphelenchus xylophilus route of transmission is the important prevention and control of current pine nematode
Strategy.The trend spread from south to north, a part of Bursaphelenchus xylophilus and Monochamus alternatus Hope distributed area have been presented in China for pine nematode
Winter temperature mostly subzero, therefore control Monochamus alternatus Hope population swollen leg bee can it is overwintering in these cold districts be to influence it
Continue the key of control effect.Field put bee experiments have shown that, the Scleroderma sichuanensis Xiao without domestication was difficult in cold district (winter
Temperature is mostly subzero) it is overwintering, this has not only seriously affected the bee to the lasting control effect of cold district Monochamus alternatus Hope population,
And greatly limit application of the bee in other cold district forest borer pest biological controls.Try to explore effectively
Domestication by low temperature method, step up the cold resistance of Scleroderma sichuanensis Xiao, obtain low temperature and resist cold strain, be to solve the bee in cold
Regional smooth overwintering key.
Summary of the invention
The object of the present invention is to provide a kind of efficient and convenient Scleroderma sichuanensis Xiao, domestication by low temperature method, this method are obtained in short-term
To Scleroderma sichuanensis Xiao cold resistance significantly increase, can get low temperature resist cold strain.
The specific technical proposal of the invention is:
A kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao, comprising the following steps:
(1) domestication kind bee selects room temperature to cultivate, the advanced age pupa of development health is as domestication material；
(2) domestication by low temperature processing 8 DEG C of 10~﹣ of ﹣ at a temperature of cultivate 7~9h make acclimation；
(3) orientation domestication first generation advanced age pupa domestication by low temperature processing postposition is cultivated under room temperature at bee eclosion, is taken into
Survive it is female carry out next-generation bee at room temperature at bee and cultivate, be developed to the pupa stage at advanced age to bee and carry out domestication by low temperature again
Processing, continuous domestication n (n≤8) generation.
The pupa that the advanced age pupa is 10~14d after pupating.
The positive effect of the present invention is embodied in:
(1) it can be used for the cold-resistant strain of acclimatization culture Scleroderma sichuanensis Xiao, meet cold district Monochamus alternatus Hope and other forestry
The needs of borer pest biological control；
(2) at a temperature of by the supercooling point of tame and docile bee, body fluid apparent freezing point and the semilethal low temperature death rate (TL50) and cold identification
The death rate significantly reduces；
(3) unaffected by the development duration of tame and docile bee and parasitized breeding ability.
Specific embodiment
Below in conjunction with specific embodiment, the present invention is described in detail, but does not limit the scope of the invention.It should be understood that
, for those of ordinary skills, can improvement or transformation based on the above description, and all these improvement and change
Changing should all belong to the protection domain of appended claims of the present invention.
The breeding method of Scleroderma sichuanensis Xiao can be with reference to the prior art or application reference number 200510020659.4 into
Row Propogation and culture.
Embodiment 1:
A kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao, comprising the following steps:
(1) domestication kind bee selects room temperature to cultivate, the advanced age pupa of development health is as domestication material；
(2) domestication by low temperature processing tames 8h in the case where 8 DEG C of ﹣；
(3) it after orientation domestication first generation advanced age pupa domestication by low temperature, is transferred to and continues culture under room temperature at bee eclosion,
It takes into survive female and carries out next-generation bee at room temperature at bee and cultivate, be developed to the pupa stage at advanced age to bee and carry out low temperature again
Acclimation, 9 generation of continuous domestication.The pupa that the advanced age pupa is 14d after pupating.
Tame effect detection
It is dead by the development duration of tame and docile bee, supercooling point, apparent freezing point, semilethal low temperature (TL50) to the 4th~9 generation respectively
The death rate and parasitized breeding ability and polypide at a temperature of rate, cold identification mainly resist cold content of material (including polypide moisture, thick rouge
The content of the small molecule alcohols such as fat and glycerol, sorbierite, mannitol and glycogen) it is detected.The result shows that domestication by low temperature is to river
The development duration and parasitized breeding ability of the swollen leg bee of sclerderm do not make significant difference (table 1 and table 5)；It is hard that domestication by low temperature can significantly reduce river
Skin swell at a temperature of the supercooling point of leg bee, apparent freezing point, semilethal low temperature (TL50) death rate and cold identification the death rate (table 2~
Table 4), greatly improve the cold-resistant substance of polypide: in conjunction with the small molecule alcohols such as water, crude fat, glycerol, sorbierite and mannitol and glycogen
Content (6~table of table 8).That is: the described domestication by low temperature method be it is feasible, the cold resistance of Scleroderma sichuanensis Xiao can be significantly improved,
It can get the cold-resistant strain of its low temperature by generation orientation domestication.
Influence of 1 domestication by low temperature of table to Scleroderma sichuanensis Xiao development duration
Note: data are 3 duplicate mean+SDs in table 1.Alphabetical difference person's difference is aobvious after data of going together in table
It writes (LSD, P < 0.05).
Influence of 2 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult supercooling point and apparent freezing point
| Handle algebra | Supercooling point | Apparent freezing point |
| Control group | -13.22±1.22b | -9.44±1.88b |
| 4 generation of domestication by low temperature in short-term | -13.96±0.4ab | -10.27±0.37ab |
| 5 generation of domestication by low temperature in short-term | -14.05±0.33ab | -10.32±0.35ab |
| 6 generation of domestication by low temperature in short-term | -14.26±0.40ab | -10.53±0.56ab |
| 7 generation of domestication by low temperature in short-term | -14.37±0.35ab | -10.78±0.81ab |
| 8 generation of domestication by low temperature in short-term | -14.46±0.36a | -10.97±0.55a |
| 9 generation of domestication by low temperature in short-term | -14.65±0.21a | -11.13±0.32a |
The influence of the death rate of 3 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult at TL50
The influence of the death rate of 4 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult at a temperature of cold identification
| Handle algebra | The death rate at a temperature of cold identification |
| Control group | 79.67±1.53c |
| 4 generation of domestication by low temperature in short-term | 62.00±4.58b |
| 5 generation of domestication by low temperature in short-term | 60.80±2.64b |
| 6 generation of domestication by low temperature in short-term | 58.29±3.12ab |
| 7 generation of domestication by low temperature in short-term | 57.79±2.65ab |
| 8 generation of domestication by low temperature in short-term | 55.67±6.66a |
| 9 generation of domestication by low temperature in short-term | 54.78±3.42a |
Influence of 5 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult fertility
| Tame algebra | Parasitic rate (%) | Egg laying amount (grain/female) |
| Control group | 83.33±1.15a | 19.47±0.89a |
| 4 generation of domestication by low temperature in short-term | 82.67±1.15a | 19.40±0.92a |
| 5 generation of domestication by low temperature in short-term | 84.12±1.89a | 19.23±1.21a |
| 6 generation of domestication by low temperature in short-term | 83.65±2.34a | 18.98±0.76a |
| 7 generation of domestication by low temperature in short-term | 82.78±1.56a | 19.57±1.02a |
| 8 generation of domestication by low temperature in short-term | 83.33±3.06a | 19.60±1.36a |
| 9 generation of domestication by low temperature in short-term | 83.94±3.67a | 19.52±1.13a |
Influence of 6 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult polypide water content
Influence of 7 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult polypide carbohydrate and crude fat content
| Tame algebra | Glycogen (ug/mg) | Total reducing sugar (ug/mg) | Crude fat (%) |
| Control group | 4.78±1.15a | 14.54±1.70a | 11.82±0.46a |
| 4 generation of domestication by low temperature in short-term | 6.64±0.38b | 15.03±0.58b | 11.99±0.35a |
| 5 generation of domestication by low temperature in short-term | 6.72±0.41b | 15.09±0.62b | 12.13±0.32a |
| 6 generation of domestication by low temperature in short-term | 6.75±0.36b | 15.21±1.53b | 12.39±0.41a |
| 7 generation of domestication by low temperature in short-term | 6.78±0.23b | 15.46±1.27b | 12.57±0.33ab |
| 8 generation of domestication by low temperature in short-term | 6.83±0.41b | 15.60±0.69b | 12.79±1.41b |
| 9 generation of domestication by low temperature in short-term | 6.91±0.32b | 15.82±1.01b | 12.81±0.35b |
Influence of 8 domestication by low temperature of table to Scleroderma sichuanensis Xiao adult polypide small molecule alcohol content
| Tame algebra | Glycerol (ug/mg) | Sorbierite (ug/mg) | Mannitol (%) |
| Control group | 3.72±0.55a | 5.09±0.58a | 0.49±0.08a |
| 4 generation of domestication by low temperature in short-term | 8.88±0.57b | 6.67±0.43b | 0.86±0.03b |
| 5 generation of domestication by low temperature in short-term | 8.91±0.45b | 7.52±0.53b | 0.89±0.04b |
| 6 generation of domestication by low temperature in short-term | 8.94±0.68b | 8.23±0.54bc | 0.94±0.03bc |
| 7 generation of domestication by low temperature in short-term | 8.97±0.65b | 8.95±0.65bc | 0.97±0.05bc |
| 8 generation of domestication by low temperature in short-term | 9.08±0.63b | 9.96±0.78c | 1.10±0.07c |
| 9 generation of domestication by low temperature in short-term | 9.14±0.58b | 10.04±0.57c | 1.12±0.06c |
Note: data are 3 duplicate mean+SDs in 2~table of table 8.It is poor with difference person alphabetical after column data in table
Different significant (LSD, P < 0.05).
Cold acclimation conditions screen Scleroderma sichuanensis Xiao in short-term, include the following steps
(1) (26 DEG C ± 1 DEG C) of room temperature cultivation of kind bee selection selection, the Scleroderma sichuanensis Xiao ovum (3d) of development health, low age
Larva (3d after hatching), high instar larvae (6d after hatching), low age pupa (7d after pupating), advanced age pupa (14d after pupating), adult (plumage
7d after change) it is used as material to be tested, material to be tested is provided by Sichuan Agricultural University's Scleroderma sichuanensis Xiao Breeding Center.
(2) low temperature exposure treatment sets 4 DEG C of ﹣, ﹣ 8 for examination Scleroderma sichuanensis Xiao ovum, larva, pupa and adult for above-mentioned respectively
DEG C, 12 DEG C of ﹣, taken out after exposure 2h, 4h, 6h, 8h at 16 DEG C of ﹣.Every processing is repeated 3 times, duplicate as follows for test worm number every time.At
Worm: 100/time；Ovum: 200 tablets/time；Larva and pupa: 200/time.With incubated at room temperature, the river without any low-temperature treatment is hard
Skin swells leg bee ovum, larva, pupa and adult as control.
(3) the culture observation after low temperature exposure will set normal culture at room temperature for examination bee after low temperature exposure treatment.To low
Ovum, larva and the pupa of temperature exposure observe its hatching, cocoon (or pupating) and emergence situation respectively, are determined with this for trying ovum, larva
With the death and survival number of pupa；To the adult of low temperature exposure, culture observes and records its dead and survival number afterwards for 24 hours.
(4) the low temperature exposure death rate compares for examination Scleroderma sichuanensis Xiao ovum, larva, pupa and the low temperature of the adult exposure death rate
It is shown in Table 9.
(5) Cold acclimation conditions screening in short-term need to generally control the death rate of material to be tested to guarantee to tame successfully
50% or so, material to be tested can be allowed to be tamed in this way, have certain survival volume after also can guarantee domestication, to be next
Wheel domestication provides material.Scleroderma sichuanensis Xiao is ectoparasite bee, and ovum and larva be attached to host's body surface, be disturbed it is easy to fall off,
Ovum and larva once fall off will the shrivelled death because cannot get nutrition supply, should not as domestication material；When being developed to pupa time
(special advanced age pupa stage), mature bee pupa can fall off from the upper nature of host, and selection is tamed smaller to the bee development impact at this time；
To the adult stage, bee ontogeny full maturity is not so good as pupa time to the sensibility of domestication by low temperature.Choosing comprehensively pros and cons, selection
The corresponding low-temperature treatment in short-term of 50% or so the death rate of advanced age pupa is as optimal acclimation conditions, it may be assumed that advanced age pupa (14d after pupating) ﹣ 8
8h is tamed at DEG C.
The exposure death rate of low temperature in short-term of 9 Scleroderma sichuanensis Xiao different developmental phases of table
Note: data are 3 duplicate mean+SDs, alphabetical difference person's significant difference after data of going together in table 9
(LSD, P < 0.05).
It should be understood that for those of ordinary skills, it can be modified or changed according to the above description,
And all these modifications and variations should all belong to the protection domain of appended claims of the present invention.
Claims (1)
1. a kind of domestication by low temperature method in short-term of Scleroderma sichuanensis Xiao, it is characterised in that the following steps are included:
(1) domestication kind bee selects room temperature to cultivate, the advanced age pupa of development health is as domestication material, and the advanced age pupa is
The pupa of 10 ~ 14d after pupating；
(2) domestication by low temperature processing set at 8 DEG C of 10 ~ ﹣ of ﹣ cultivate 7 ~ 9h make acclimation；
(3) orientation domestication will put back to culture under room temperature and take and survive down at bee eclosion after first generation advanced age pupa domestication by low temperature
Come it is female carries out next-generation bee cultivation at room temperature at bee, be developed to the pupa stage at advanced age to bee and carry out at domestication by low temperature again
It manages, continuous domestication n generation, wherein n≤8.
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