CN103329854A - Soybean aphid homogenized artificial breeding method - Google Patents
Soybean aphid homogenized artificial breeding method Download PDFInfo
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- CN103329854A CN103329854A CN201310287480XA CN201310287480A CN103329854A CN 103329854 A CN103329854 A CN 103329854A CN 201310287480X A CN201310287480X A CN 201310287480XA CN 201310287480 A CN201310287480 A CN 201310287480A CN 103329854 A CN103329854 A CN 103329854A
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Abstract
The invention discloses a soybean aphid homogenized artificial breeding method which comprises a host plant homogenized processing technology, a soybean aphid homogenized selection cultivation technology, a soybean aphid homogenized inoculation and separation cultivation technology, and soybean aphid homogenized collection technology. The method can achieve soybean aphid large-size low-cost and standardized production, soybean aphids can be provided at any time for related scientific researches and study works like biological control pesticide new product development.
Description
Technical field
The present invention relates to a kind of artificial cultivating method of homogenization of small insects, particularly possess the artificial cultivating method of homogenization of the soybean aphid of sucking mouth parts.
Background technology
Soybean aphid (Aphis glycines Matsmura) is commonly called as " aphid ", belongs to Homoptera aphid section, is one of primary pest of soybean.Soybean aphid often is gathered in the tender stem of soybean, the tender leaf back side, sucks juice, causes blade to crispatura, growth retardation, and early leaf fall, root system development is bad, and branch and the quantity that bears pods reduce, and 100-grain weight descends, and output reduces, even whole strain death.In addition, soybean aphid still is the main communication media of soybean mosaic virus.
Soybean aphid has become the important worldwide pest of extensive concern.Therefore, be badly in need of the new soybean varieties of the anti-aphid of seed selection, the breeding expert has also carried out the anti-aphid molecular breeding work of soybean one after another both at home and abroad.
Yet the soybean aphid of various places has different biotypes, and the host is very narrow, and overwintering host only limits to the several plant of Rhamnus, and the summer host only limits to each cultivation and wild soybean kind, and breeding work only limits to obtain for 6 to October the worm source every year.Development test needs the aphid in homogeneous stage; The aphid source that can obtain in the anniversary.Also do not develop at present specially and culture aphid at the artificial feed anniversary of soybean aphid.
Therefore, be badly in need of the problem of propagating artificially of the aphid in solution homogeneous stage at present, and with lower cost, breed the soybean aphid source of different ecological type fast, for the further exploitation of the anti-aphid breeding of soybean and the anti-novel biopesticide that eliminates aphis provides safeguard and supports.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of soybean aphid homogenization artificial cultivating method is provided.The present invention can provide the homogenization soybean aphid of sufficient amount efficiently, easily at short notice, and the research work relevant with soybean aphid can be carried out smoothly.
A kind of soybean aphid homogenization artificial cultivating method provided by the invention comprises the steps:
1) homogenization of host plant is handled;
2) homogenization of soybean aphid is selected to cultivate;
3) homogenization of soybean aphid inoculation is cultivated with isolation;
4) homogenization of soybean aphid is collected.
The homogenization of described step 1) host plant is handled, concrete operations are: organic substrate is added clear water stir behind high-temperature sterilization, plantation soybean sense aphid kind, condition of culture is illumination 15h-18h/ days, 26 ± 1 ℃, bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, begin to inoculate soybean aphid.
Preferably, soybean sense aphid kind is " east farming 47 ", " closing rich 47 " or " Williams82 ".
Described step 2) homogenization of soybean aphid is selected to cultivate, concrete operations are: several the aptery soybean aphids of maturation are seeded on the sense aphid soybean leaves of cultured in vitro, excised leaf encases the glass culture dish that the petiole end places sterilization with moistening absorbent cotton, cultivate the breeding nymph, remove all aptery aphids after 24 hours, only select wherein 1 age in days aphid, cultivated 72-76 hour, the homogeneous nymph of gathering breeding carries out inoculation experiments.Aphid raises up seed approximately needs 3 day time, and used afterwards aphid all is the offspring of this aphid breeding, guarantees the homogeneity of aphid.
Preferably, condition of culture is illumination: dark=14:10, and intensity of illumination 300 μ molm-2s-1, the control illumination temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.
The homogenization of described step 3) soybean aphid is inoculated and is isolated cultivation, and concrete operations are: select 1 age in days nymph of cultivation to brush gently to the plant leaf of step 1) gained with moistening writing brush homogenization and breed.
Preferably, with transparent cylindrical glass cartridge postvaccinal soybean seedling cover is got up, tighten with 150-180 purpose screen cloth on the glass infuser top, be placed in the Turnover Box that fills water, and at the cystosepiment of a high 2-3cm of the bottom of basin pad, keep the water surface in the Turnover Box just not have the bottom of basin, and add water in good time, be placed on the breeding of carrying out soybean aphid in the phytotron.More preferably, condition of culture is illumination 16h-18h/ days, 25 ± 1 ℃.The glass infuser height is 40-50cm, and diameter is 15-18cm.
The homogenization collection technique of described step 4) soybean aphid, concrete operations are: cultured soybean aphid is taken off together with blade, be placed on a side of culture dish, opposite side at culture dish is placed the 3-5cm * cloth of 2cm yellow background and fresh soybean leaves, utilize aphid to the taxis of yellow, aphid is lured to new soybean leaves experiment and research after carrying out.
The present invention includes following advantage:
1, the homogenization treatment technology of host plant can be selected suitable host, and soybean aphid can breed on host plant fast in a large number, has shortened the breeding cycle, has increased aphid output.
2, the homogenization of soybean aphid selects culture technique can make the standardization breeding of aphid, is the aphid of handling through homogenization because just begun used aphid, so the aphid offspring of breeding can guarantee the selectivity of its homogeneity and virulence.
3. the homogenization of soybean aphid inoculation and isolation culture technique, incubation is in the cloche of isolation always, can not cause the heterogeneity of population because of mixing between the aphid individuality, thereby follow-up experimental result is produced interference.No season dependence, can annual carry out soybean aphid fast and a large amount of breeding.
4, the homogenization collection process technology of soybean aphid utilizes aphid to the taxis of yellow, aphid is lured to new soybean leaves can more accurately be identified the soybean varieties resistance.
The artificial fecundation method of soybean aphid provided by the present invention can have been realized standardization and the scale breeding of soybean aphid; and cost is low; simple to operate, for the exploitation of the anti-aphid breeding work of follow-up soybean, novel biological pesticide and the basic scientific research work relevant with soybean aphid provide support and ensure.
Description of drawings
Fig. 1: through the soybean aphid that separation obtains, carry out the homogenization breeding.
Fig. 2: soybean aphid is seeded in after homogenization is handled on the soybean sense aphid kind and breeds.
Fig. 3: the soybean aphid of the homogenization of expanding propagation.
Fig. 4: the soybean aphid that homogenization is collected.
Embodiment
Embodiment 1
(1) organic substrate being added proper amount of clear water behind high-temperature sterilization stirs.Selecting soybean varieties is " east farming 47 ", and condition of culture is illumination 16h/ days, 26 ℃.Bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, can supply the inoculation soybean aphid.
(2) several the aptery soybean aphids of maturation are seeded on the sense aphid soybean leaves of cultured in vitro, excised leaf encases the glass culture dish that the petiole end places sterilization with moistening absorbent cotton.Preferably, condition of culture is illumination: dark=14:10, intensity of illumination 300 μ molm
-2S
-1, the control illumination temperature is 24 ℃, dark temperature is 22 ℃.Remove all aptery aphids after 24 hours, only select the wherein nymph of a 1-d-old, cultivated 72 hours, the homogeneous nymph of gathering breeding carries out inoculation experiments.
(3) selecting the soybean aphid of cultivation to brush gently to blade with moistening writing brush homogenization breeds.Preferably, with transparent cylindrical glass cartridge (high 45cm, diameter 15cm) postvaccinal soybean seedling cover is got up, tighten with 150 purpose screen clothes on the glass infuser top, be placed in the Turnover Box that fills water, and at the cystosepiment of a high 2--3cm of the bottom of basin pad, keep the water surface in the Turnover Box just not have the bottom of basin, and add water in good time, be placed on the breeding of carrying out soybean aphid in the phytotron.More preferably, condition of culture is illumination 16h/ days, 24 ℃.
(4) cultured soybean aphid is taken off together with blade, be placed on a side of culture dish, preferably, opposite side at culture dish is placed the cloth of 3cm*2cm yellow background and fresh soybean leaves, utilize aphid to the taxis of yellow, aphid is lured to new soybean leaves experiment and research after carrying out.
Through the homogenization artificial cultivating method of soybean aphid provided by the present invention, aphides gene type stable and consistent after measured, reproduction rate improves 10%, has realized standardization and the scale breeding of soybean aphid.
Embodiment 2
(1) organic substrate being added proper amount of clear water behind high-temperature sterilization stirs.Select soybean varieties to be " closing rich 47 " sense aphid kind, condition of culture is illumination 18h//sky, 27 ℃.Bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, can supply the inoculation soybean aphid.
(2) several the aptery soybean aphids of maturation are seeded on the sense aphid soybean leaves of cultured in vitro, excised leaf encases the glass culture dish that the petiole end places sterilization with moistening absorbent cotton.Preferably, condition of culture is illumination: dark=14:10, intensity of illumination 300 μ molm
-2S
-1, the control illumination temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.Remove all aptery aphids after 24 hours, only select the wherein nymph of a 1-d-old, cultivated 74 hours, the homogeneous nymph of gathering breeding carries out inoculation experiments.
(3) selecting the soybean aphid of cultivation to brush gently to blade with moistening writing brush homogenization breeds.Preferably, with transparent cylindrical glass cartridge (high 50cm, diameter 18cm) postvaccinal soybean seedling cover is got up, tighten with 180 purpose screen clothes on the glass infuser top, be placed in the Turnover Box that fills water, and at the cystosepiment of a high 3cm of the bottom of basin pad, keep the water surface in the Turnover Box just not have the bottom of basin, and add water in good time, be placed on the breeding of carrying out soybean aphid in the phytotron.More preferably, condition of culture is illumination 18h/ days, 25 ℃.
(4) cultured soybean aphid is taken off together with blade, be placed on a side of culture dish, preferably, opposite side at culture dish is placed the cloth of 5cm*2cm yellow background and fresh soybean leaves, utilize aphid to the taxis of yellow, aphid is lured to new soybean leaves experiment and research after carrying out.
Through the homogenization artificial cultivating method of soybean aphid provided by the present invention, aphides gene type stable and consistent after measured, reproduction rate improves 10%, has realized standardization and the scale breeding of soybean aphid.
Embodiment 3
(1) organic substrate being added proper amount of clear water behind high-temperature sterilization stirs.Selecting soybean varieties is " Williams82 " sense aphid kind, and condition of culture is illumination 17h//sky, 24 ℃.Bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, can supply the inoculation soybean aphid.
(2) several the aptery soybean aphids of maturation are seeded on the sense aphid soybean leaves of cultured in vitro, excised leaf encases the glass culture dish that the petiole end places sterilization with moistening absorbent cotton.Preferably, condition of culture is illumination: dark=14:10, intensity of illumination 300 μ molm
-2S
-1, the control illumination temperature is 23 ℃, dark temperature is 21 ℃.Remove all aptery aphids after 24 hours, only select the wherein nymph of a 1-d-old, cultivated 76 hours, the homogeneous nymph of gathering breeding carries out inoculation experiments.
(3) selecting the soybean aphid of cultivation to brush gently to blade with moistening writing brush homogenization breeds.Preferably, with transparent cylindrical glass cartridge (high 45cm, diameter 16cm) postvaccinal soybean seedling cover is got up, tighten with 165 purpose screen clothes on the glass infuser top, be placed in the Turnover Box that fills water, and at the cystosepiment of a high 2.5cm of the bottom of basin pad, keep the water surface in the Turnover Box just not have the bottom of basin, and add water in good time, be placed on the breeding of carrying out soybean aphid in the phytotron.More preferably, condition of culture is illumination 17h/ days, 23 ℃.
(4) cultured soybean aphid is taken off together with blade, be placed on a side of culture dish, preferably, opposite side at culture dish is placed the cloth of 4cm*2cm yellow background and fresh soybean leaves, utilize aphid to the taxis of yellow, aphid is lured to new soybean leaves experiment and research after carrying out.
Through the homogenization artificial cultivating method of soybean aphid provided by the present invention, aphides gene type stable and consistent after measured, reproduction rate improves 10%, has realized standardization and the scale breeding of soybean aphid.
More than describe preferred embodiment of the present invention in detail.The ordinary skill that should be appreciated that this area need not creative work and just can design according to the present invention make many modifications and variations.Therefore, all technical staff in the art all should be in the determined protection domain by claims under this invention's idea on the basis of existing technology by the available technical scheme of logical analysis, reasoning, or a limited experiment.
Claims (6)
1. a soybean aphid homogenization artificial cultivating method comprises the steps:
1) homogenization of host plant is handled;
2) homogenization of soybean aphid is selected to cultivate;
3) homogenization of soybean aphid inoculation is cultivated with isolation;
4) homogenization of soybean aphid is collected;
The homogenization of described step 1) host plant is handled, concrete operations are: organic substrate is added clear water stir behind high-temperature sterilization, plantation soybean sense aphid kind, condition of culture is illumination 15h-18h/ days, 26 ± 1 ℃, bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, begin to inoculate soybean aphid;
Described step 2) homogenization of soybean aphid is selected to cultivate, concrete operations are: several the aptery soybean aphids of maturation are seeded on the sense aphid soybean leaves of cultured in vitro, excised leaf encases the glass culture dish that the petiole end places sterilization with moistening absorbent cotton, cultivate the breeding nymph, remove all aptery aphids after 24 hours, and only select 1 an age in days nymph to continue to cultivate, and to cultivate 72-76 hour, the homogeneous nymph of gathering breeding carries out inoculation experiments;
The homogenization of described step 3) soybean aphid is inoculated and is isolated cultivation, and concrete operations are: select 1 age in days nymph of cultivation to brush gently to the plant leaf of step 1) gained with moistening writing brush homogenization and breed;
The homogenization of described step 4) soybean aphid is collected, concrete operations are: cultured soybean aphid is taken off together with blade, be placed on a side of culture dish, opposite side at culture dish is placed the 3-5cm * cloth of 2cm yellow background and fresh soybean leaves, utilize aphid to the taxis of yellow, aphid is lured to new soybean leaves experiment and research after carrying out.
2. the method for claim 1, wherein in the step 1), described soybean sense aphid kind is " east farming 47 ", " closing rich 47 " or " Williams82 ".
3. the method for claim 1, wherein step 2) in, condition of culture is illumination: dark=14:10, and intensity of illumination 300 μ molm-2s-1, the control illumination temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.
4. the method for claim 1, wherein in the step 3), with transparent cylindrical glass cartridge postvaccinal soybean seedling cover is got up, tighten with 150-180 purpose screen cloth on the glass infuser top, be placed in the Turnover Box that fills water, and at the cystosepiment of a high 2-3cm of the bottom of basin pad, keep the water surface in the Turnover Box just not have the bottom of basin, and add water in good time, be placed on the breeding of carrying out soybean aphid in the phytotron.
5. method as claimed in claim 4, wherein in the step 3), condition of culture is illumination 16h-18h/ days, 25 ± 1 ℃.
6. method as claimed in claim 5, wherein in the step 3), the glass infuser height is 40-50cm, diameter is 15-18cm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103875607A (en) * | 2014-03-14 | 2014-06-25 | 上海交通大学 | Method for identifying soybean aphid physiological races |
| CN107306897A (en) * | 2017-08-17 | 2017-11-03 | 上海交通大学 | A kind of device for limiting aphid scope of activities and the method for pinpointing captive breeding aphid |
| CN109247303A (en) * | 2018-11-16 | 2019-01-22 | 湖北省农业科学院粮食作物研究所 | Utilize the method for water planting semen viciae fabae seedling mass propagation bean aphid |
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| CN101601375A (en) * | 2008-06-12 | 2009-12-16 | 中国农业科学院植物保护研究所 | A kind of artificial fecundation method of aphid |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103875607A (en) * | 2014-03-14 | 2014-06-25 | 上海交通大学 | Method for identifying soybean aphid physiological races |
| CN103875607B (en) * | 2014-03-14 | 2016-05-04 | 上海交通大学 | The authentication method of a kind of soybean aphid biological strain |
| CN107306897A (en) * | 2017-08-17 | 2017-11-03 | 上海交通大学 | A kind of device for limiting aphid scope of activities and the method for pinpointing captive breeding aphid |
| CN109247303A (en) * | 2018-11-16 | 2019-01-22 | 湖北省农业科学院粮食作物研究所 | Utilize the method for water planting semen viciae fabae seedling mass propagation bean aphid |
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