CN113575518A - Method for insect film toxicant feeding and application thereof - Google Patents
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Abstract
本发明属于麦蚜生理学与植物病理学技术领域，具体涉及一种刺吸式昆虫薄膜饲毒的方法及其应用。该方法包括以下步骤：(1)将塑料膜均匀拉薄后罩在两端开口的容器上端；(2)滴加饲毒液于薄膜上；(3)将塑料膜均匀拉薄后罩在饲毒液上；(4)将所述容器下端开口处倾斜向上，将所述昆虫通过下端开口放入所述容器中；(5)将装了昆虫的容器的下端开口朝下置于吸水纸上，置于平面上进行饲毒饲养。该方法操作简单方便，而且有效提高了昆虫吸食食物或者毒液的效率，为进一步研究昆虫与植株之间的传毒效率提供了便利。
The invention belongs to the technical field of wheat aphid physiology and plant pathology, and particularly relates to a method for feeding poison with a piercing-sucking insect film and its application. The method includes the following steps: (1) uniformly thinning a plastic film and then covering the upper end of the container with two open ends; (2) dripping feeding venom on the film; (3) uniformly stretching the plastic film and then covering the feeding venom (4) the lower opening of the container is tilted upward, and the insects are put into the container through the lower opening; (5) the lower opening of the container containing the insects is placed on the absorbent paper, and the Poisonous feeding was carried out on a flat surface. The method is simple and convenient to operate, and effectively improves the efficiency of insects sucking food or venom, which provides convenience for further research on the efficiency of transmission between insects and plants.
Description
Technical Field
The invention belongs to the technical field of wheat aphid physiology and phytopathology, and particularly relates to a method for feeding poison to a piercing-sucking insect film and application thereof.
Background
Barley Yellow Dwarf Virus (BYDVs) can infect more than 150 kinds of cereal crops such as wheat, Barley, rice, oat and the like and grassy weeds. Barley, wheat, rye, triticale, oat, wild oat, sweet corn, brome, green bristlegrass, tiger tail, rabdosia rubescens, broussonetia papyrifera, haynaldia villosa, silver grass, golden green bristlegrass and the like can be infected in China, wherein self-growing wheat seedlings, tiger tail, wild oat, golden green bristlegrass, small brow grass and the like are overwintering hosts of BYDVs. The yellow dwarf disease of wheat crop caused by the virus is one of the most serious virus diseases of cereal crop. The common symptoms after the plant is infected with diseases are dwarfing, yellowing of leaves, reddening, leaf rolling and the like. The main symptom of wheat is that the leaves are chlorosis, generally spread from the leaf apex or leaf edge to the base, the middle and lower parts of the leaves are longitudinal lines with yellow-green intervals, and internodes are shortened and dwarfed. BYDV occurs to varying degrees in wheat production areas in Europe, America, Asia, oceania, etc., and causes significant losses. The disease is caused in winter wheat, spring wheat, winter spring wheat and the like in northwest, north China, northeast, east China, southwest and the like, and is mainly prevalent in winter spring wheat in north China. In 1987, yellow dwarf causes the loss of wheat in Gansu province and Shanxi province to be more than 5 hundred million kilograms. In 1998, disasters are prevalent in large areas, and the occurrence range of the disasters is spread over multiple provinces (autonomous regions) such as Shaanxi, Gansu, Ningxia, inner Mongolia, Shanxi and Hebei.
BYDVs are transmitted in a recurrent non-proliferative manner using aphids as mediators. When BYDVs move in the body of aphids and the aphids eat plants with viruses, the viruses enter the intestinal tract along with phloem juice through an oral needle and pass through the intestinal membrane of the rear intestine to enter the blood cavity under the receptor-mediated endocytosis-efflux action. The virus is distributed in the haemolymph gland and, when it reaches the accessory salivary gland, it passes through the accessory salivary gland with the accessory salivary gland basal plate in another receptor-mediated endocytosis-efflux manner and finally enters the salivary compartment. When aphids feed on another plant, the virus is secreted with saliva to invade its phloem. BYDVs were divided into 5 lines depending on aphid mediator transmission specificity: PAV, RPV, SGV, MAV, RMV. PAV is transmitted by non-specialization such as sitobion avenae and sitobion avenae; the RPV is transmitted by the specialization of the gloiopeltis aurantiaca; SGV is transmitted by the idiosyncratic transmission of the binary wheat aphids; MAV is transmitted by myzus avenae specialization; RMV was transmitted exclusively by corn aphids. Later, BYDVs were divided into two subgroups based on serological relationships and cytopathological differences: subgroup I and subgroup II. Subgroup I comprises PAV, SGV and MAV strains, subgroup II comprises RMV and RPV strains, and GPV is a specific strain type in China.
Whether aphid is transmitted or not and the transmission efficiency have important influence on the occurrence and prevalence of yellow dwarf disease. On one hand, the virus epidemic prediction can be carried out by knowing the difference of the virus transmission efficiency between aphid species and within the aphid species, which shows that the aphid clone line with obvious genetic difference is likely to have regional and regional epidemics, and the seasonal growth of the genotype with high or low virus transmission efficiency in a certain region influences the virus epidemic. On the other hand, comparison of the transmission efficiency among genotypes and hybridization of genotypes with larger differences help to find the transmission-related genes and determine the molecular sites of aphid virus interaction. However, due to the specific characteristic of the BYDVs transmitted by the wheat aphids, the virus transmission efficiency has certain difference; and different plants or different reproduction modes of wheat aphid clone lines, the virus transmission efficiency is greatly different, so that the virus disease monitoring and early warning are very difficult. Particularly, when the virus transmission efficiency is researched, the defects of complicated process, low efficiency of sucking food viruses by aphids and the like exist in the aphid virus feeding technology, and the research difficulty of spreading yellow dwarf by aphids is increased.
The present document, simple method of aphid membrane feeding poison of von Chongchuan, plant protection, discloses a simple method of aphid membrane feeding poison, which is characterized by that the PARAFILM of paraffin preparation is lightly pulled into developed membrane which is as thin as possible by hand and covered on the container mouth, and the membrane is adhered on the surface, then a semi-transparent plastic tube with diameter of 0.5cm is used to cut into rings with height of 0.8-1 mm, and used as membrane type supporting ring, three supporting rings are placed on the membrane by using tweezers, then the aphid is dropped on the surface of the membrane by using injector, finally, a membrane is covered on the liquid surface, and due to the action of supporting ring, an inner chamber with edible liquid is stored between two membranes, and the edible liquid is fully distributed on the surface of the membrane. The thin film toxicant feeding container is inverted, and then aphids are picked into the container, so that the aphids are all on the inner membrane of the thin film chamber. Because the film has elasticity, the film chamber causes certain osmotic pressure, and the aphid can smoothly take the food. And (3) when the aphids start to prick and suck, turning the thin-film toxin feeding container upside down, enabling the bottle mouth to be upward, and plugging the middle hole opening by using a cotton ball or sticking the cotton ball to paper. The method has complex and inconvenient process, and needs to judge whether the aphids start to prick or not, certain subjective judgment factors influence the operation process, and the toxicity transmission rate of the aphids cannot be ensured.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for insect membrane feeding, which can effectively improve the efficiency of feeding a feeding solution to small insects with piercing-sucking mouthparts.
The method comprises the following steps:
(1) after the plastic film is uniformly thinned, covering the upper end of the container with openings at two ends;
(2) dropping feeding venom on the film;
(3) uniformly thinning the plastic film and covering the plastic film on the venom;
(4) inclining the opening at the lower end of the container upwards, and putting the insects into the container through the opening at the lower end;
(5) placing the container with insect on the water-absorbing paper with its lower end opening facing downwards, and placing on a plane for feeding;
the insects are small insects with piercing-sucking mouthparts.
Preferably, the insects are 2-year old insects for 3-4 days.
Further, the container may be any regular or irregular shaped container.
Preferably, the container is a glass cylinder.
Preferably, the diameter of the glass cylinder is 3-4cm, more preferably 3.5 cm.
Preferably, the volume of the venom is 0.1-0.3ml, more preferably 0.2 ml.
Preferably, the number of 2-year-old insects of 3-4 days is 30-40.
Preferably, in the step (3), after the plastic film is uniformly thinned and covered on the venom, air bubbles between the films are removed.
Preferably, the opening at the lower end of the container is inclined upwards at an angle of 30-60 °, preferably 45 °.
Further, the preparation method of the venom comprises the following steps: squeezing equal amount of the plants with toxin with water to obtain juice, centrifuging, and adding sucrose. It should be noted that the venom herein can be replaced with other food or substance for small insects with piercing-sucking mouthparts.
Preferably, the insect is wheat aphid and the fed virus is barley yellow dwarf virus.
More preferably, the myzus avenae is myzus avenae, and the barley yellow dwarf virus is a Chinese taiwanese disease strain or a Belgiwen disease strain. The propagation efficiency of the ophiocordyceps avenae population in different areas to the diseased plant in different areas is different.
The invention further provides a method for detecting the virus transmission efficiency of insects, which is characterized in that insects are enabled to suck sap containing venom based on a thin-film virus feeding mode, and then insects with viruses gnaw plants for virus transmission, so that the accuracy of the virus transmission efficiency is improved. Then, the efficiency of transmitting the barley yellow dwarf virus by different geographical populations or different kinds of wheat aphids is identified by a method for transmitting the barley yellow dwarf virus by feeding the wheat aphids by an indoor thin film method, so that the occurrence condition of local virus diseases can be accurately predicted, and a basis is provided for mastering the epidemic rule of BYDVs.
The detection method comprises the following steps:
(1) obtaining the insect with toxicity by using the insect film toxicity feeding method in any one of the previous methods;
(2) inoculating the virus to the plant seedlings in the two-leaf stage;
(3) after the plant seedlings are inoculated with viruses, the insects are removed;
(4) identifying the virus-carrying condition of the plant seedlings;
(5) obtaining the insect virus transmission efficiency according to the number of virus-carrying plant strains/the number of all detected plant strains multiplied by 100 percent;
in the step (4), DAS-ELISA is adopted to identify the virus carrying condition of the plant seedlings; if the OD405nm value of the tested plant is more than 3 times of the negative control OD405nm value, the plant successfully carries the virus, and the plant is a virus-carrying plant; the negative control is a non-inoculated virus plant.
In particular, the DAS-ELISA is an experimental method routinely used by those skilled in the art.
Further, the method of removing the insects may be a medicinal repellent or a physical repellent, in order to stop the insects from sucking.
Further, in the step (2), the time for inoculating the virus to the plant seedlings in the two-leaf stage is more than or equal to 48 hours.
The invention has the beneficial effects that
The insect film feeding method provided by the invention is simple and convenient to operate, effectively improves the efficiency of the insects for sucking food or venom, and provides convenience for further researching the efficiency of the transmission of the toxin between the insects and plants.
The method for detecting the efficiency of the insect to transmit the virus can evaluate the efficiency of the barley yellow dwarf virus transmitted by the wheat aphid, can timely and accurately predict the occurrence condition of the local virus disease by evaluating the occurrence trend of the barley yellow dwarf virus disease, can grasp the epidemic rule of BYDVs, and has important significance for the occurrence of the barley yellow dwarf virus disease, the trial prediction of a hazard area, the large-area production layout and the scientific medication.
Drawings
FIG. 1 is a schematic view of the detection process of the virus transmission efficiency of wheat aphids.
Detailed Description
The examples are given for the purpose of better illustration of the invention, but the invention is not limited to the examples. Therefore, those skilled in the art should make insubstantial modifications and adaptations to the embodiments of the present invention in light of the above teachings and remain within the scope of the invention.
In the embodiment of the present invention, the detection of barley yellow dwarf virus transmitted by sitobion avenae is performed with reference to the schematic flow chart shown in fig. 1, and the specific detection method is as follows:
(1) selecting a plant variety to be detected;
(2) feeding the 2 nd aphid with artificial feed film for 48 h;
(3) inoculating aphids to plant seedlings at two leaf stages, inoculating three heads to each seedling for 30 seedlings, and setting feeding nontoxic artificial feed as a control;
(4) spraying a medicament on the inoculated plant seedlings after 3 days to thoroughly kill aphids;
(5) after 15 days, symptoms were observed and the diseased plants were counted using DAS-ELISA identification.
Each population was run under the same conditions for the same procedure, repeated three times for each population.
In the embodiment of the invention, the membrane toxicity feeding method comprises the following steps:
(1) uniformly thinning the sealing film and covering the sealing film on the upper end of a glass cylinder with the diameter of 3.5cm and two open ends;
(2) 0.2ml of the feeding venom is dripped on the membrane,
(3) after being uniformly thinned, the sealing film covers the feeding venom, so that liquid drops are prevented from flowing out, and air bubbles between the film and the culture dish cover are removed;
(4) the opening at the lower end of the glass column is inclined upwards at an angle of about 45 degrees, and 30-40 myzus persicae of 2-year-old avenae for 3-4 days are gently placed into the glass column through the opening at the lower end;
(5) the lower end of the glass column added with the aphids is downwards opened and is placed on the absorbent paper, and the glass column is placed on a flat table top for feeding.
In the implementation of the invention, the preparation method of the venom comprises the following steps:
(1) putting fresh leaves of the plants with the toxicity into a transparent sealed sample bag, and weighing;
(2) adding equal weight of water into the sample bag;
(3) squeezing plant juice with a squeezer;
(4) transferring the juice to a 2ml centrifuge tube by using a pipette;
(5) centrifugation at 13.2rpm for 2 minutes;
(6) sucking 1ml of supernatant and placing the supernatant into a new centrifuge tube;
(7) adding 0.2g of sucrose into a centrifuge tube;
(8) shaking and mixing evenly.
In the embodiment of the invention, the method for detecting the test sample by using a double-antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) comprises the following steps:
(1) the virus-carrying wheat seedlings are used as a positive control of an ELISA experiment, the wheat without inoculated virus strains is used as a negative control, and PBS buffer solution is used as a blank control. Determining the OD at 405nm on an enzyme-linked reader;
(2) coating antibody: and simultaneously setting a blank control, a positive control and a negative control according to the number of the samples to be detected. Adding virus antiserum (1: l000 diluted in coating buffer) to wells of an enzyme-linked plate at 200. mu.l per well, covering and moisturizing, and incubating at 37 ℃ for 4 hours;
(3) preparing a sample: weighing 0.1g of sample tissue, putting the sample tissue into a centrifuge tube, adding l.9ml of coating buffer solution, squeezing juice on FastPrep, and taking supernatant for later use;
(4) washing the plate: pouring out the anti-coating solution on the enzyme-linked plate, washing the plate for 3 times by using PBS-Tween buffer solution, each time for 5 minutes, and draining;
(5) binding to antigen: add 200. mu.l sample supernatant per well, incubate overnight at 4 ℃ with moisture;
(6) washing the plate: the method is the same as (4);
(7) binding an enzyme-labeled antibody; add 200. mu.l of IgG labeled with alkaline phosphatase diluted 1000 times with ligation buffer (PBS-Tween containing 2% PVP) per well, 200. mu.l per well, cover and moisturize, incubate for 4 hours at 37 ℃;
(8) washing the plate: the method is the same as (4) or (6);
(9) color development: adding a newly prepared substrate solution, reacting for 60mins at room temperature, wherein each well is 200 mul;
(10) and (3) determination: after observing the color reaction, determining the OD value at 405nm on an enzyme labeling instrument;
(11) virus-carrying strain identification: if the OD value of the tested plant is more than 3 times of the negative control OD value, the virus transmission of the plant is successful and the plant is a virus strain;
(12) the myzus avenae transmission rate is equal to the number of virus strains/the number of all tested strains is 100%.
Example 1 detection of the transmission efficiency of barley yellow dwarf virus (Chinese Taiyuan disease strain) by Myzus avenae of different geographical populations
The propagation efficiency of the 14 geographical populations of central aphids in China to propagate the Chinese Taiyuan BYDV-PAV strain is detected by taking the wheat variety Beijing 837 as a detection plant, and the detection result is shown in Table 1.
TABLE 1 spreading efficiency of different geographical groups on barley yellow dwarf (Chinese Taiyuan plant)
Note:athe letters in the same column indicate significant differences at the level of 0.05 (df-13, MS-8.219, F-23.5, P)<0.0001,HSD test)。cThe number of aphids without toxicity.
As can be seen from Table 1, the transmission efficiency of BYDV-PAV Chinese Taiyuan strains transmitted by Myzus avenae in different geographical populations in China has certain difference, the average virus transmission efficiency of the tested populations is 35.08%, and the variation range of the virus transmission efficiency is from 23.55% to 56.18%. The virus-transmitting efficiency of the Shanxi Taiyuan population is 56.18 +/-5.22 percent, while the virus-transmitting efficiency of the Henan Deng state population is only 23.55 +/-1.36 percent. The highest virus transmission efficiency is 2.4 times higher than the lowest virus transmission efficiency. HSD detection shows that the virus transfer efficiency between the middle and lower Yangtze river population (Anhui Hefei, Hubei Dangjiang Kou, Henan Deng Zhou, Hubei Ziyang, Jiangsu Zhenjiang, Jiangsu Yancheng and Sichuan Jiang oil)) and the Yunnan red river population is not different; there was no difference between the northern population (Shandong Taian and Shanxi Taiyuan population) and the northwest population (Qinghai Xining, Shaanxi Bao chicken and Xinjiang Shihe Zi population). In addition, it can be seen that the virus transmission efficiency of the north and northwest populations is higher than that of the middle and lower reaches of Yangtze river.
Example 2 detection of the efficiency of transmission of barley yellow dwarf virus (Belleville Luwensis disease Strain) by Myzus avenae in different geographical populations
The propagation efficiency of 14 geographical populations of Myzus avenae in China to propagate the Wewen BYDV-PAV strain Belgium was examined using the wheat variety Beijing 837 as the test plant, and the test results are shown in Table 2.
TABLE 2 efficiency of transmission of barley yellow dwarf (Belgium Luwen disease strain) by different geographical groups
Note:athe letters in the same column indicate significant differences at the level of 0.05 (df-13, MS-8.219, F-23.5, P)<0.0001,HSD test)。cThe number of aphids without toxicity.
As can be seen from Table 2, BYDV-PAV Belgium strains can be transmitted by the wheat aphids in China, which shows that the method can successfully carry out the virus transmission of the wheat aphids of different geographical populations to the cross-regional virus strains. The average virus transmission efficiency of the tested population is 42.06%, and the virus transmission efficiency ranges from 24.42% to 66.67%. The virus transfer efficiency of the Shanxi Taiyuan population is 66.67 +/-3.84%, while the virus transfer efficiency of the Henan Deng state population is only 24.42 +/-2.21%. The highest virus transmission efficiency is 3 times higher than the lowest virus transmission efficiency. HSD detection shows that the virus transfer efficiency between the middle and lower Yangtze river population (Anhui Hefei, Hubei Dangjiang Kou, Henan Deng Zhou, Hubei Ziyang, Jiangsu Zhenjiang, Jiangsu Yancheng and Sichuan Jiang oil)) and the Yunnan red river population is not different; there was no difference between the northern population (Shandong Taian and Shanxi Taiyuan population) and the northwest population (Qinghai Xining, Shaanxi Bao chicken and Xinjiang Shihe Zi population). In addition, it can be seen that the virus transmission efficiency of the north and northwest populations is higher than that of the middle and lower reaches of Yangtze river.
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (10)
1. A method for insect membrane detoxification, said method comprising the steps of:
(1) after the plastic film is uniformly thinned, covering the upper end of the container with openings at two ends;
(2) dropping feeding venom on the film;
(3) uniformly thinning the plastic film and covering the plastic film on the venom;
(4) inclining the opening at the lower end of the container upwards, and putting the insects into the container through the opening at the lower end;
(5) placing the container with insect on the water-absorbing paper with its lower end opening facing downwards, and placing on a plane for feeding;
the insects are small insects with piercing-sucking mouthparts.
2. The method of claim 1, wherein the insect is a 2-year old insect from 3 to 4 days.
3. The method of claim 1, wherein the container is a glass cylinder.
4. The method of claim 3, wherein said glass cylinder has a diameter of 3-4cm and said volume of said venom is 0.1-0.3 ml.
5. The method of claim 1, wherein the number of 2-year-old insects from 3 to 4 days is 30 to 40.
6. The method according to claim 1, wherein in the step (3), the plastic film is uniformly thinned and covered with the venom, and then air bubbles are removed from the film.
7. The method of claim 1, wherein in step (4), the lower opening of the container is inclined upward at an angle of 30 ° to 60 °.
8. The method of claim 1, wherein the venom is prepared by: squeezing equal amount of the plants with toxin with water to obtain juice, centrifuging, and adding sucrose.
9. The method according to any one of claims 1 to 8, wherein the insect is Myzus persicae and the fed virus is barley yellow dwarf virus.
10. A method for detecting the efficiency of transmission of a virus by an insect, said method comprising the steps of:
(1) obtaining a virulent insect using the method of any one of claims 1-8;
(2) inoculating the virus to the plant seedlings in the two-leaf stage;
(3) after the plant seedlings are inoculated with viruses, the insects are removed;
(4) identifying the virus-carrying condition of the plant seedlings;
(5) obtaining the insect virus transmission efficiency according to the number of virus-carrying plant strains/the number of all detected plant strains multiplied by 100 percent;
in the step (4), DAS-ELISA is adopted to identify the virus carrying condition of the plant seedlings; if the OD of the tested plant405nmValue of>3-fold negative control OD405nmIf so, the virus is successfully transmitted to the plant, and the plant is a virus-carrying plant; the negative control is a non-inoculated virus plant.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5899168A (en) * | 1995-03-15 | 1999-05-04 | The United States Of America, As Represented By The Secretary Of Agriculture | Synthetic diet for rearing the hymenopterous ectoparasitoid, catolaccus grandis |
| CN102754623A (en) * | 2012-07-24 | 2012-10-31 | 浙江大学 | Method for acquiring ricedrawfvirus (RDV) by utilizing nephotettix cincticeps under artificial conditions and application of RDV |
| CN102870743A (en) * | 2012-10-12 | 2013-01-16 | 江苏省农业科学院 | Method for preserving rice black-streaked dwarf virus indoor living bodies |
| CN107047485A (en) * | 2017-04-27 | 2017-08-18 | 福建省农业科学院茶叶研究所 | A kind of method of Cicadellidae insect indoor feeding and bionical raising box |
| CN206413611U (en) * | 2016-11-16 | 2017-08-18 | 华南农业大学 | A kind of piercing sucking insect feeding device |
| CN110396503A (en) * | 2019-07-22 | 2019-11-01 | 中国农业大学 | A method for acquiring brassica yellowing virus of the genus Potato leafroll virus by using peach aphid |
| CN209965007U (en) * | 2018-10-09 | 2020-01-21 | 中国农业科学院植物保护研究所 | Device for biological evaluation of piercing-sucking mouthpart insects |
| JP2020065548A (en) * | 2018-10-19 | 2020-04-30 | 国立大学法人東京農工大学 | Arthropod feeding device and feeding method |
| CN211379324U (en) * | 2019-12-28 | 2020-09-01 | 赣南师范大学 | A kind of citrus psyllid artificial feeding observation cup |
-
2021
- 2021-07-27 CN CN202110849418.XA patent/CN113575518A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5899168A (en) * | 1995-03-15 | 1999-05-04 | The United States Of America, As Represented By The Secretary Of Agriculture | Synthetic diet for rearing the hymenopterous ectoparasitoid, catolaccus grandis |
| CN102754623A (en) * | 2012-07-24 | 2012-10-31 | 浙江大学 | Method for acquiring ricedrawfvirus (RDV) by utilizing nephotettix cincticeps under artificial conditions and application of RDV |
| CN102870743A (en) * | 2012-10-12 | 2013-01-16 | 江苏省农业科学院 | Method for preserving rice black-streaked dwarf virus indoor living bodies |
| CN206413611U (en) * | 2016-11-16 | 2017-08-18 | 华南农业大学 | A kind of piercing sucking insect feeding device |
| CN107047485A (en) * | 2017-04-27 | 2017-08-18 | 福建省农业科学院茶叶研究所 | A kind of method of Cicadellidae insect indoor feeding and bionical raising box |
| CN209965007U (en) * | 2018-10-09 | 2020-01-21 | 中国农业科学院植物保护研究所 | Device for biological evaluation of piercing-sucking mouthpart insects |
| JP2020065548A (en) * | 2018-10-19 | 2020-04-30 | 国立大学法人東京農工大学 | Arthropod feeding device and feeding method |
| CN110396503A (en) * | 2019-07-22 | 2019-11-01 | 中国农业大学 | A method for acquiring brassica yellowing virus of the genus Potato leafroll virus by using peach aphid |
| CN211379324U (en) * | 2019-12-28 | 2020-09-01 | 赣南师范大学 | A kind of citrus psyllid artificial feeding observation cup |
Non-Patent Citations (2)
| Title |
|---|
| 于文娟 等: "抗生素对麦长管蚜传毒效率的影响", 《"创新驱动与现代植保"——中国植物保护学会第十一次全国会员代表大会暨2013年学术年会论文集》 * |
| 汪东风 等: "《食品质量与安全检测技术（第三版）》", 陕西科学技术出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114158525A (en) * | 2021-11-11 | 2022-03-11 | 西南大学 | A method for rapid and precise drug selection in the field of Panonychus citrus |
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