CN103329854B - Soybean aphid homogenized artificial breeding method - Google Patents
Soybean aphid homogenized artificial breeding method Download PDFInfo
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- CN103329854B CN103329854B CN201310287480.XA CN201310287480A CN103329854B CN 103329854 B CN103329854 B CN 103329854B CN 201310287480 A CN201310287480 A CN 201310287480A CN 103329854 B CN103329854 B CN 103329854B
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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 is often 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 declines, yield reducation, even whole strain is dead.In addition, the main communication media of soybean aphid or 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, 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, host is very narrow, and overwintering host only limits to the several plant of Rhamnus, and summer host only limits to each cultivation and wild soybean kind, and breeding work only limits to obtain worm source every year 6 to October.Development test needs the aphid of equal one-phase; The aphid source that can obtain in the anniversary.Also do not develop at present the special cultivation of the artificial feed anniversary for soybean aphid aphid.
Therefore, be badly in need of at present the problem of propagating artificially of the aphid of the equal one-phase of solution, and with lower cost, breed fast the soybean aphid source of different ecological type, for the further exploitation of the breeding of Soybean Resistance aphid and the anti-novel biopesticide eliminating aphis provides safeguard and supports.
Summary of the invention
The object of the invention is to for the deficiencies in the prior art, a kind of soybean aphid homogenization artificial cultivating method is provided.The present invention can be efficiently, the homogenization soybean aphid of sufficient amount is provided easily at short notice, and the research work relevant to 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 processed;
2) homogenization of soybean aphid is selected to cultivate;
3) homogenization inoculation and the isolate forster of soybean aphid;
4) homogenization of soybean aphid is collected.
Described step 1) homogenization of host plant is processed, concrete operations are: organic substrate is added after high-temperature sterilization to clear water and stir, 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, start to inoculate soybean aphid.
Preferably, soybean sense aphid kind is " eastern agriculture 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 with moistening absorbent cotton the glass culture dish that petiole end is placed in sterilizing, cultivate breeding nymph, after 24 hours, remove all aptery aphids, only select a wherein 1 age in days aphid, cultivate 72-76 hour, the homogeneous nymph that gathers breeding carries out inoculation experiments.Aphid raises up seed approximately needs 3 day time, and aphid used is all the offspring of this aphid breeding afterwards, guarantees the homogeneity of aphid.
Preferably, condition of culture is, illumination: dark=14:10, intensity of illumination 300 μ molm
-2s
-1, controlled light temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.
Described step 3) homogenization inoculation and the isolate forster of soybean aphid, concrete operations are: the 1 age in days nymph that homogenization select is cultivated is brushed to step 1 gently with moistening writing brush) breed on the plant leaf of gained.
Preferably, by transparent cylindrical glass cartridge, postvaccinal soybean seedling cover is got up, tighten with 150-180 object screen cloth on 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 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 phytotron.More preferably, condition of culture is illumination 16h-18h/ days, 25 ± 1 ℃.Glass infuser height is 40-50cm, and diameter is 15-18cm.
Described step 4) the homogenization collection technique of 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 cloth of 3-5cm * 2cm yellow background and fresh soybean leaves, utilize aphid to yellow taxis, aphid is lured to new soybean leaves to 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 is amount reproduction on host plant fast, 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 processing through homogenization, so the aphid offspring of breeding can guarantee the selectivity of its homogeneity and virulence because just started aphid used.
3. the homogenization inoculation of soybean aphid and isolate forster technology, incubation in the cloche in isolation, can not cause because of mixing between aphid individuality the heterogeneity of population, thereby follow-up experimental result is produced and disturbed always.Without dependence in season, can annual carry out the quick and amount reproduction of soybean aphid.
4, the homogenization collection process technology of soybean aphid, utilizes aphid to yellow taxis, and aphid is lured to precise Identification soybean varieties resistance more on new soybean leaves.
The artificial fecundation method of soybean aphid provided by the present invention can have been realized standardization and the biological control of soybean aphid; and cost is low; simple to operate, for the exploitation of follow-up Soybean Resistance aphid breeding work, Novel biocontrol agricultural chemicals and the basic scientific research work relevant to soybean aphid provide support and ensure.
Accompanying drawing explanation
Fig. 1: the soybean aphid being isolated to, carries out homogenization breeding.
Fig. 2: soybean aphid is seeded in soybean sense aphid kind and breeds after homogenization is processed.
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 after high-temperature sterilization to proper amount of clear water stirs.Selecting soybean varieties is " eastern agriculture 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 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 with moistening absorbent cotton the glass culture dish that petiole end is placed in sterilizing.Preferably, condition of culture is, illumination: dark=14:10, intensity of illumination 300 μ molm
-2s
-1, controlled light temperature is 24 ℃, dark temperature is 22 ℃.After 24 hours, remove all aptery aphids, only select the wherein nymph of a 1-d-old, cultivate 72 hours, the homogeneous nymph that gathers breeding carries out inoculation experiments.
(3) select the soybean aphid of cultivating to brush gently to blade and breed with moistening writing brush homogenization.Preferably, with transparent cylindrical glass cartridge (high 45cm, diameter 15cm) postvaccinal soybean seedling cover is got up, tighten with 150 object screen clothes on 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 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 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 yellow taxis, aphid is lured to new soybean leaves to 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 biological control of soybean aphid.
Embodiment 2
(1) organic substrate being added after high-temperature sterilization to proper amount of clear water stirs.Select soybean varieties for " closing rich 47 " sense aphid kind, condition of culture is illumination 18h/ days, 27 ℃.Bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, can supply 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 with moistening absorbent cotton the glass culture dish that petiole end is placed in sterilizing.Preferably, condition of culture is, illumination: dark=14:10, intensity of illumination 300 μ molm
-2s
-1, controlled light temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.After 24 hours, remove all aptery aphids, only select the wherein nymph of a 1-d-old, cultivate 74 hours, the homogeneous nymph that gathers breeding carries out inoculation experiments.
(3) select the soybean aphid of cultivating to brush gently to blade and breed with moistening writing brush homogenization.Preferably, with transparent cylindrical glass cartridge (high 50cm, diameter 18cm) postvaccinal soybean seedling cover is got up, tighten with 180 object screen clothes on 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 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 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 yellow taxis, aphid is lured to new soybean leaves to 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 biological control of soybean aphid.
Embodiment 3
(1) organic substrate being added after high-temperature sterilization to proper amount of clear water stirs.Selecting soybean varieties is " Williams82 " sense aphid kind, and condition of culture is illumination 17h/ days, 24 ℃.Bean seedlings grow to V1 vegetative period, i.e. first trifoliolate leaf phase, can supply 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 with moistening absorbent cotton the glass culture dish that petiole end is placed in sterilizing.Preferably, condition of culture is, illumination: dark=14:10, intensity of illumination 300 μ molm
-2s
-1, controlled light temperature is 23 ℃, dark temperature is 21 ℃.After 24 hours, remove all aptery aphids, only select the wherein nymph of a 1-d-old, cultivate 76 hours, the homogeneous nymph that gathers breeding carries out inoculation experiments.
(3) select the soybean aphid of cultivating to brush gently to blade and breed with moistening writing brush homogenization.Preferably, with transparent cylindrical glass cartridge (high 45cm, diameter 16cm) postvaccinal soybean seedling cover is got up, tighten with 165 object screen clothes on 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 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 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 yellow taxis, aphid is lured to new soybean leaves to 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 biological control of soybean aphid.
More than describe preferred embodiment of the present invention in detail.The ordinary skill that should be appreciated that this area just can design according to the present invention be made many modifications and variations without creative work.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 processed;
2) homogenization of soybean aphid is selected to cultivate;
3) homogenization inoculation and the isolate forster of soybean aphid;
4) homogenization of soybean aphid is collected;
Described step 1) homogenization of host plant is processed, concrete operations are: organic substrate is added after high-temperature sterilization to clear water and stir, 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, start 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 with moistening absorbent cotton the glass culture dish that petiole end is placed in sterilizing, cultivate breeding nymph, after 24 hours, remove all aptery aphids, and only select a 1 age in days nymph to continue to cultivate, and cultivate 72-76 hour, the homogeneous nymph that gathers breeding carries out inoculation experiments;
Described step 3) homogenization inoculation and the isolate forster of soybean aphid, concrete operations are: the 1 age in days nymph that homogenization select is cultivated is brushed to step 1 gently with moistening writing brush) breed on the plant leaf of gained;
Described step 4) homogenization of 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 cloth of 3-5cm * 2cm yellow background and fresh soybean leaves, utilize aphid to yellow taxis, aphid is lured to new soybean leaves to experiment and research after carrying out.
2. the method for claim 1, wherein step 1) in, described soybean sense aphid kind is, " eastern agriculture 47 ", " closing rich 47 " or " Williams82 ".
3. the method for claim 1, wherein step 2) in, condition of culture is, illumination: dark=14:10, intensity of illumination 300 μ molm
-2s
-1, controlled light temperature is 24 ± 1 ℃, dark temperature is 22 ± 1 ℃.
4. the method for claim 1, step 3 wherein) in, by transparent cylindrical glass cartridge, postvaccinal soybean seedling cover is got up, tighten with 150-180 object screen cloth on 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 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 phytotron.
5. method as claimed in claim 4, wherein step 3) in, condition of culture is illumination 16h-18h/ days, 25 ± 1 ℃.
6. method as claimed in claim 5, wherein step 3) in, glass infuser height is 40-50cm, diameter is 15-18cm.
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| 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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| CN101477098A (en) * | 2009-01-08 | 2009-07-08 | 上海交通大学 | Method for detecting aphid resistance of soybean by using soybean natural extract |
| 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 |
| CN102648703A (en) * | 2011-02-28 | 2012-08-29 | 汪鹏丞 | Artificial aphid breeding and honeydew preparation technique |
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Patent Citations (3)
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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 |
| CN101477098A (en) * | 2009-01-08 | 2009-07-08 | 上海交通大学 | Method for detecting aphid resistance of soybean by using soybean natural extract |
| CN102648703A (en) * | 2011-02-28 | 2012-08-29 | 汪鹏丞 | Artificial aphid breeding and honeydew preparation technique |
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| 徐蕾等.温度对大豆蚜生长发育和繁殖的影响.《中国油料作物学报》.2011,第33卷(第2期),第189-192页. * |
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