CN110835655A - Method for preventing and treating virus disease of trilobate cotton leaf curl - Google Patents
Method for preventing and treating virus disease of trilobate cotton leaf curl Download PDFInfo
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Abstract
The invention relates to a method for preventing and treating a leaf curl virus disease of hibiscus syriacus cotton; the method comprises the steps of 1) dividing the disease condition grading standard of the virus disease of the hibiscus syriacus; 2) performing PCR detection on the Multan cotton leaf curl virus, and judging whether the Multan cotton leaf curl virus is infected; 3) and (3) comprehensively preventing and treating the hibiscus syriacus in the next year according to the grading standard of the step 1 and the contamination confirmation of the step 2. The method has the beneficial effects that the comprehensive control of the hibiscus syriacus relates to the growth and development stage of the hibiscus syriacus, in particular to the agricultural control measures in the rooting stage and the transplanting stage; four toxicity test experiments are carried out, and thorough experiments find that the high-efficiency cyhalothrin and the abamectin have very outstanding effects on the control of bemisia tabaci which is a target organism of a pathogenic hibiscus syriacus plant. The method and the device can be widely popularized in the industry, and can also provide theoretical basis for prevention and treatment methods of other plants and viruses thereof.
Description
Technical Field
The invention belongs to the technical field of a prevention and treatment method of a hibiscus syriacus cotton leaf curl virus, and particularly relates to a specific detection primer and an β satellite molecule primer of the hibiscus syriacus cotton leaf curl virus, and further comprises a comprehensive prevention and treatment method of the hibiscus syriacus virus.
Background
Hibiscus rosa-sinense, also known as Hibiscus sinensis, Carthamus tinctorius, etc., belongs to the genus Hibiscus of the family Malvaceae. The hibiscus syriacus is mainly distributed in parts of south China, southwest China and east China, and is an important garden ornamental plant in the south China. The Multan Cotton leaf curl virus (CLCuMuV) is an important virus causing the leaf curl disease of the hibiscus syriacus, and after a plant is infected with diseases, the symptoms of the plant are that the whole leaf vein is obviously enlarged (figure 1A), the leaves are curled upwards (figure 1B), the leaf vein is enlarged seriously to form leaf ears, finally, the plant is weak in growth vigor, little or no flowering exists, the leaves are yellowed and died finally (figure 1C). Under natural conditions, the hibiscus leaf curl disease is mainly transmitted by bemisia tabaci in a lasting way, and additionally, cuttage propagation and virus-carrying seedlings are also transmitted artificially in a long distance but cannot be transmitted by mechanical friction and seed virus carrying. In addition to the fact that the Multan cotton leaf curl virus infects the hibiscus syriacus, the Multan cotton leaf curl virus can also infect host plants such as cotton, okra, flower of pending boll, corchorus fasciatus and the like and cause leaf curl disease symptoms in China. The hibiscus leaf curl disease caused by CLCuMuV infection is discovered in Guangdong province of China for the first time in 2006, and then, in Guangxi, Hainan, Fujian, Yunnan, Jiangsu, Xinjiang and other places successively, in recent years, the hibiscus is taken as an important landscaping plant and a family potted flower and is widely transported across the country, garden nurseries and bases for mass production of hibiscus seedlings are provided in Guangdong Guangxi and Fujian areas, and the potential risk of diffusion and outbreak of CLCuMuV to cotton areas of China is greatly increased by transportation of virus-carrying hibiscus seedlings and diffusion of virus-carrying media bemisia tabaci. On the premise of accurately identifying the pathogenic species and the classification status thereof, dynamic tracking monitoring of field occurrence is carried out aiming at pathogenic viruses and virus vectors [ bemisia tabaci (figure 1D) ], it is clear that the major reason for long-distance spreading of the hibiscus leaf curl virus is transregional distribution of disease-sensitive hosts, the major reason for short-distance natural spreading is spread by the virus-carrying vectors and virus-carrying hosts, and the key point of disease control is explored aiming at the problem that the virus is passively carried and spread along with virus-carrying host plants or by means of virus-carrying medium insects.
Disclosure of Invention
The invention aims to solve the defects of the problems and provides a method for preventing and treating the leaf curl virus disease of hibiscus syriacus cotton.
The invention is realized by adopting the following technical scheme.
The invention discloses a specific primer of a Multan cotton leaf curl virus, which is a primer comprising CL-F: CAGGAAGCAGGAAAATACGAGA, respectively; CL-R: TGGCAGTCCAACACAAAATACG are provided.
An β satellite molecule specific primer of the Multan cotton leaf curl virus, the β satellite molecule specific primer beta-F: AAGTCGAATGGAACGTGAATGT and beta-R: GGAGACCAAAAGAGGAGAGAGA are provided.
The method for detecting the Multan cotton leaf curl virus by using the primers is PCR detection, and the PCR conditions are as follows: 25 μ LPCR reaction: 2 muL (10 ng-20 ng) of DNA template, 12.5 muL of PCR Mixer, 1 muL of each upstream primer and downstream primer of 10 mumol/L and 8.5 muL of sterilized water; and (3) PCR reaction conditions: pre-denaturation at 94 ℃ for 4 min; denaturation at 94 ℃ for 45s, annealing at 52 ℃ for 45s, extension at 52 ℃ for 1min, 35 cycles, and extension at 72 ℃ for 10 min.
The method for comprehensively preventing and treating the hibiscus syriacus by using the detection method comprises the steps of 1) dividing disease condition grading standard of hibiscus syriacus virus; 2) performing PCR detection on the Multan cotton leaf curl virus, and judging whether the Multan cotton leaf curl virus is infected and whether the infection is serious; 3) comprehensive control of hibiscus syriacus was confirmed in the next year according to the grading criteria of step 1 and infection of step 2.
The grading standard of the disease condition of the virus of the hibiscus syriacus is as follows,
the comprehensive control method comprises the following steps: rooting and potting time: in the south, most roots after 20 days, and the seedlings can grow into pots after growing for more than one month. In cold places, heating is adopted to keep the temperature at 20-25 ℃, the seedlings are inserted into a sand basin, water is sprayed for 1-3 times a day, roots grow after more than 40 days, and the seedlings are planted in the basin after 60 days; transplanting: after the seedlings are transplanted to root, the seedlings are potted; covering the limbers at the bottom of the pot with tiles, filling base fertilizer in the bottom layer, filling mixed soil, planting seedlings, watering thoroughly, watering 1 time a day, and shading; after seedling delaying, the seedlings are placed in a place with sufficient sunlight, and pots are changed after 2-3 years of cultivation, wherein the pot changing time is carried out in the end of spring and in the early summer.
Further, the comprehensive control of the invention comprises: field management: outdoor cultivation and indoor cultivation, shading in the early stage after cuttage, keeping the relative humidity at 85% -95%, controlling the temperature at 18-25 ℃, sufficiently illuminating, and topdressing once in half a month. In northern cold area, the temperature is controlled at 15-22 deg.C after 10 months.
Further, the comprehensive control of the invention comprises: and (3) monitoring the quantity of the bemisia tabaci by using a yellow hanging plate in the seedling stage and the adult stage. When the bemisia tabaci is low in density, yellow pest sticking plates can be hung in the shed or in the field, and the hanging height is the same as that of the plant, and each 20m pest sticking plate is hung2 Hang 1 piece.
Further, the comprehensive control of the invention comprises: MEAM1 cryptophyte and asiai ii 7 cryptophyte were killed using high potency cyhalothrin.
Further, the comprehensive control of the invention comprises: the abamectin is used for killing the hidden seeds of MEAM1 and the hidden seeds of Asia II 7.
The method has the beneficial effects that the steps are related, the β satellite molecule specific primers of the Multan cotton leaf curl virus are designed on the basis of firstly setting the disease condition grading standard of the Hibiscus syriacus virus to carry out specific detection on the virus, the plant infection virus can be confirmed according to strips on an electrophoresis chart, and the comprehensive control on the Hibiscus syriacus in the next year is carried out by combining the grading standard of the step 1 and the virus confirmation of the step 2.
The invention is further explained below with reference to the drawings and the detailed description.
Drawings
FIG. 1A shows the green and swollen symptoms of the leaf vein of the leaf curl of hibiscus syriacus.
FIG. 1B is a picture showing the upward curling symptom of leaf of Arthropoda cinnabarina.
FIG. 1C is a graph showing that whole plant died due to Arthropoda cinnabarina leaf curl disease.
FIG. 1D is a photograph of Bemisia tabaci, a transmission mediator of the virus of Hibiscus syriacus leaf curl.
FIG. 2 is the electrophoresis chart of the detection of the xylem cotton leaf curl virus of the hibiscus syriacus leaf curl disease plant, No. 1-6 are 6 hibiscus syriacus samples with disease onset in different places respectively.
FIG. 3 shows an β satellite molecular detection electrophoresis chart of a hibiscus syriacus leaf curl disease plant, wherein Nos. 1-6 are 6 hibiscus syriacus samples with onset at different sites.
FIG. 4 is a detection electrophoresis of a Hibiscus syriacus plant suspected of leaf curl, and Nos. 1-6 are 6 Hibiscus syriacus plants in different locations, respectively.
Detailed Description
Disease condition grading standard of hibiscus syriacus virus disease
CLCuMuV detection of hibiscus syriacus leaves
The total DNA of the hibiscus syriacus plant leaves is extracted by adopting a plant cell tissue genome kit EF111-12 and Beijing holotype gold biotechnology limited company, the specific operation is carried out according to the kit specification, and PCR detection is respectively carried out by adopting a CLCuMuV specific primer CL-F, CL-R and a β satellite molecule specific primer beta-F, beta-R (table 1).
25 μ LPCR reaction: 2. mu.L (. about.20 ng) of DNA template, 12.5. mu.L of PCR Mixer, 1. mu.L of each of the upstream and downstream primers at 10. mu. mol/L, and 8.5. mu.L of sterilized water.
And (3) PCR reaction conditions: pre-denaturation at 94 ℃ for 4 min; denaturation at 94 ℃ for 45s, annealing at 52 ℃ for 45s, extension at 52 ℃ for 1min, 35 cycles, and extension at 72 ℃ for 10 min.
After the PCR reaction, 10. mu.L of the LPCR product was electrophoresed on a 1% agarose gel.
TABLE 1 primers in PCR analysis
Prevention and control technology for hippeastrum leaf curl disease
The prevention and control of the hibiscus leaf curl disease are developed by taking the guidance idea of 'prevention as the main part and comprehensive prevention and control'.
1. Strengthen the quarantine of the seedling and ensure the safety of the production of the seedling
(1) Cutting propagation is mostly adopted in production, outdoor cultivation is combined with early spring pruning, cutting is carried out in 5-6 months, but if indoor cutting is carried out, cutting is generally combined with early spring pruning in 1-2 months.
(2) Land selection and preparation: selecting weak acid soil which faces the sun, has fertile soil and good loosening and drainage.
(3) Seedling selection: selecting a healthy and healthy hibiscus syriacus adult plant as a stock plant of the cutting. Selecting annual half-woody robust branches, cutting the branches to have the length of 8-l0cm, removing the lower end leaves completely, cutting half of the upper leaves to leave 3-4 leaves, and cutting the branches into a nutrient medium or a sand pot.
(4) Seedling management: 1/3 are inserted into the coarse sand and then are watered thoroughly, and the plant-row spacing is 4cm multiplied by 4 cm. In order to maintain the air temperature, a film is covered.
(5) Rooting and potting time: in the south, most roots after 20 days, and the seedlings can grow into pots after growing for more than one month. In cold places, the temperature is kept at 20-25 ℃ by heating, the seedlings are inserted into sand pots, water is sprayed for 1-3 times a day, roots grow after more than 40 days, the seedlings are planted in the pots after 60 days, and the survival rate is 60%.
(6) And (3) other daily management: and when the bemisia tabaci exists, timely preventing and removing the chemical agent. When plants with obvious diseases are found, the plants are immediately destroyed, and the batch of seedlings are subjected to insecticidal protection.
(7) Cleaning a seedling planting environment: cleaning withered branches and leaves, removing peripheral weeds, bringing the treatment to outdoor centralized treatment, and controlling mass propagation of a virus-transmitting medium, namely bemisia tabaci and a potential virus source carried by the same.
2. Strengthen the production safety of the nursery stock in the adult plant period
(1) Transplanting: and (5) transplanting the seedlings into a pot after the seedlings are transplanted to root. Covering the water holes on the bottom of the pot with tiles, filling base fertilizer on the bottom layer, filling mixed soil, planting seedlings, watering thoroughly, watering 1 time a day, and shading. After seedling delaying, the seedlings are placed in a place with sufficient sunlight, and pots are changed after 2-3 years of cultivation, wherein the pot changing time is carried out in the end of spring and in the early summer.
(2) Field management: outdoor cultivation and indoor cultivation, shading in the early stage after cuttage, keeping the relative humidity at 85% -95%, controlling the temperature at 18-25 ℃, sufficiently illuminating, and topdressing once in half a month. In northern cold area, the greenhouse is controlled at 15-22 deg.C at the end of 10 months, and ventilation, less fertilizer application or culture application is performed.
3. Control of the transmitted virus mediator bemisia tabaci
And (3) in the seedling stage and the adult stage, yellow hanging plates are used for monitoring the quantity of the bemisia tabaci. When the bemisia tabaci is low in density, yellow pest sticking plates can be hung in the shed or in the field, and the hanging height is the same as that of the plant, and each 20m pest sticking plate is hung2 Hang 1 piece, in time clear up the change. When the density is high, the registered insecticide and the compound agent thereof can be selected for spraying prevention and control. The purpose of disease and insect prevention is achieved.
The bemisia tabaci on the hibiscus syriacus is an invasive cryptophyte MEAM1 and a local cryptophyte Asia II 7, the local cryptophyte Asia II 7 can transmit the virus of the hibiscus syriacus leaf curl, and in order to clarify the control effect of the common bemisia tabaci medicament on the bemisia tabaci on the hibiscus syriacus, the invention carries out control tests on the two bemisia tabaci seeds. The selected medicaments are as follows: 50% Acetamiprid (Acetamiprid) missible oil, 20% Imidacloprid (Imidacloprid) soluble solution, 1% high-efficiency cyhalothrin (Lambda-cyhalothrin) emulsion in water, 1.8% avermectin (Avermectins) missible oil, and the sensitivity test of the pesticide is carried out at the temperature of 26 +/-2 ℃, the humidity of 80% +/-5%, the light cycle L: d ═ 14: 10 in a greenhouse.
Each medicament is provided with 5 concentrations and 1 group of blank control, the hidden species of the MEAM1 is 5000 times of the liquid of each medicament, and the hidden species of the AsiaI 7 is 9000 times of the liquid of each medicament, respectively. The method is characterized in that an agar moisturizing and leaf soaking method is adopted, according to the improvement of the method of Feng et al (2009), 5mL of 1.5% agar liquid is added at the bottom of a flat-bottom glass tube (with the diameter of 28mm multiplied by the height of 12cm), after the agar is naturally cooled and solidified, cotton leaves are beaten into round leaves with the diameter of 26mm by using a puncher, the leaves are immersed into liquid medicine with various concentrations for 15s, taken out and aired, then the back of the leaves is upward, the leaves are flatly placed on the agar in the flat-bottom glass tube, and the agar is treated with clear water to serve as a control. Each pipe is connected with 30-40 adult bemisia tabaci and sealed by a 120-mesh gauze. The glass tube was inverted at a temperature of 26. + -. 2 ℃ with a photoperiod L: d ═ 14: in a 10 light incubator, the mortality of each treatment was examined after 48 h.
Indoor toxicity test results show (Table 2) that for the MEAM1 cryptic species, the high-efficiency cyhalothrin and the abamectin LC502.163mg/L and 2.867mg/L respectively, imidacloprid LC5041.136mg/L of acetamiprid LC5084.013mg/L, which is 39 times of high-efficiency cyhalothrin.
For Asia II 7 cryptophyte, high-efficiency cyhalothrin and abamectin LC501.624mg/L and 1.672mg/L respectively, imidacloprid LC5027.136mg/L of acetamiprid LC5057.163mg/L, which is 35 times of high-efficiency cyhalothrin. The sensitivity of the MEAM1 and Asia II 7 cryptophytes to 4 agents is basically the same, and the virulence is high-efficiency cyhalothrin>Abamectin and its preparation method>Imidacloprid>Acetamiprid, whichThe control effect of the first two medicaments is better.
TABLE 2 virulence assays for the four insecticides against MEAM1 and Asia II 7 Bemisia tabaci
Therefore, for preventing and treating the medium AsiaI II 7 bemisia tabaci of the virus disease of the hibiscus syriacus leaf curl, 5000-time liquid of high-efficiency cyhalothrin or 5000-time liquid of abamectin is recommended to be selected. When the pesticide is applied, the pesticide is uniformly sprayed on the front and back of the leaf for 2-3 times at an interval of 5-7 days.
The above description is only a part of specific embodiments of the present invention (since the formula of the present invention belongs to the numerical range, the embodiments are not exhaustive, and the protection scope of the present invention is subject to the numerical range and other technical point ranges), and the detailed contents or common knowledge known in the schemes are not described too much. It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by means of equivalent substitution or equivalent transformation for those skilled in the art are within the protection scope of the present invention. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
<110> institute for plant protection of academy of agricultural sciences of Guangdong province
<120> method for preventing and treating cotton leaf curl virus disease of hibiscus syriacus
<160>4
<210>1
<211>22
<212>DNA
<213> Artificial sequence
<400>1
CAGGAAGCAGGAAAATACGAGA
<210>2
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<213> Artificial sequence
<400>2
TGGCAGTCCAACACAAAATACG
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<213> Artificial sequence
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AAGTCGAATGGAACGTGAATGT
<210>4
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<213> Artificial sequence
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GGAGACCAAAAGAGGAGAGAGA
Claims (10)
1. The specific primer of the Multan cotton leaf curl virus is characterized by being CL-F: CAGGAAGCAGGAAAATACGAGA, respectively; CL-R: TGGCAGTCCAACACAAAATACG are provided.
2. An β satellite molecule specific primer of the Multan cotton leaf curl virus is characterized in that the β satellite molecule specific primer beta-F: AAGTCGAATGGAACGTGAATGT, beta-R:
GGAGACCAAAAGAGGAGAGAGA。
3. a method for detecting a multan cotton leaf curl virus using the primers of claim 1 or 2, characterized in that the method is a PCR detection, and the PCR conditions are: 25 μ LPCR reaction: 2 muL of DNA template 10 ng-20 ng, PCR mixer12.5 muL, 1 muL of upstream and downstream primers of 10 mumol/L, 8.5 muL of sterilized water; and (3) PCR reaction conditions: pre-denaturation at 94 ℃ for 4 min; denaturation at 94 ℃ for 45s, annealing at 52 ℃ for 45s, extension at 52 ℃ for 1min, 35 cycles, and extension at 72 ℃ for 10 min.
4. The method for the comprehensive control of hibiscus syriacus by using the method as claimed in claim 3, characterized by comprising the steps of 1) dividing disease grading standard of hibiscus syriacus virus disease; 2) carrying out PCR detection on the Multan cotton leaf curl virus, and further verifying the leaves infected with the Multan cotton leaf curl virus; 3) and (3) comprehensively preventing and treating the hibiscus syriacus according to the grading standard of the step 1 and the contamination confirmation of the step 2.
5. The method for the comprehensive control of hibiscus syriacus according to claim 4, wherein the disease grading standard of hibiscus syriacus virus disease is as follows,
6. the method for the integrated control of hibiscus syriacus as claimed in claim 4, wherein the integrated control comprises: rooting and potting time: in the south, most roots after 20 days, and the seedlings can grow into pots after growing for more than one month. In cold places, heating is adopted to keep the temperature at 20-25 ℃, the seedlings are inserted into a sand basin, water is sprayed for 1-3 times a day, roots grow after more than 40 days, and the seedlings are planted in the basin after 60 days; transplanting: after the seedlings are transplanted to root, the seedlings are potted; covering the limbers at the bottom of the pot with tiles, filling base fertilizer in the bottom layer, filling mixed soil, planting seedlings, watering thoroughly, watering 1 time a day, and shading; after seedling delaying, the seedlings are placed in a place with sufficient sunlight, and pots are changed after 2-3 years of cultivation, wherein the pot changing time is carried out in the end of spring and in the early summer.
7. The method for the integrated control of hibiscus syriacus as claimed in claim 4, wherein the integrated control comprises: field management: outdoor cultivation and indoor cultivation, shading in the early stage after cuttage, keeping the relative humidity at 85% -95%, controlling the temperature at 18-25 ℃, sufficiently illuminating, and topdressing once in half a month. In northern cold area, the temperature is controlled at 15-22 deg.C after 10 months.
8. The method for the integrated control of hibiscus syriacus as claimed in claim 4, wherein the integrated control comprises: and (3) monitoring the quantity of the bemisia tabaci by using a yellow hanging plate in the seedling stage and the adult stage. When the bemisia tabaci is low in density, yellow pest sticking plates can be hung in the shed or in the field, and the hanging height is the same as that of the plant, and each 20m pest sticking plate is hung2Hang 1 piece.
9. The method for the integrated control of hibiscus syriacus as claimed in claim 4, wherein the integrated control comprises: MEAM1 cryptophyte and asiai ii 7 cryptophyte were killed using high potency cyhalothrin.
10. The method for the integrated control of hibiscus syriacus as claimed in claim 4, wherein the integrated control comprises: the abamectin is used for killing the hidden seeds of MEAM1 and the hidden seeds of Asia II 7.
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| CN112048573A (en) * | 2020-09-28 | 2020-12-08 | 广东省农业科学院植物保护研究所 | RPA primer and kit for detecting cotton leaf curl virus, detection method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102939880A (en) * | 2012-11-27 | 2013-02-27 | 福建省农业科学院植物保护研究所 | Standardized planting method for black plum |
| CN103238435A (en) * | 2013-05-09 | 2013-08-14 | 新疆农业科学院经济作物研究所 | Method for constructing cotton population structure of jujube and cotton intercropping drip irrigation cotton field in Xinjiang |
| CN103960092A (en) * | 2014-05-06 | 2014-08-06 | 云南省烟草公司玉溪市公司 | Three-dimensional biological control method for plant diseases and insect pests of tobacco planting |
| CN104031988A (en) * | 2014-05-07 | 2014-09-10 | 中国农业科学院烟草研究所 | Identification method for physiological races of erysiphe cichoracearum DC. |
| CN109730083A (en) * | 2019-02-02 | 2019-05-10 | 湖南省植物保护研究所 | Bemisia tabaci approach behavior adjusting control agent, the application of insecticide and insecticide, the application of desinsection plate and desinsection plate |
| CN109811040A (en) * | 2019-02-21 | 2019-05-28 | 北京农业智能装备技术研究中心 | A kind of germ quantitative detecting method based on PMA dyeing |
-
2019
- 2019-12-10 CN CN201911261771.5A patent/CN110835655A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102939880A (en) * | 2012-11-27 | 2013-02-27 | 福建省农业科学院植物保护研究所 | Standardized planting method for black plum |
| CN103238435A (en) * | 2013-05-09 | 2013-08-14 | 新疆农业科学院经济作物研究所 | Method for constructing cotton population structure of jujube and cotton intercropping drip irrigation cotton field in Xinjiang |
| CN103960092A (en) * | 2014-05-06 | 2014-08-06 | 云南省烟草公司玉溪市公司 | Three-dimensional biological control method for plant diseases and insect pests of tobacco planting |
| CN104031988A (en) * | 2014-05-07 | 2014-09-10 | 中国农业科学院烟草研究所 | Identification method for physiological races of erysiphe cichoracearum DC. |
| CN109730083A (en) * | 2019-02-02 | 2019-05-10 | 湖南省植物保护研究所 | Bemisia tabaci approach behavior adjusting control agent, the application of insecticide and insecticide, the application of desinsection plate and desinsection plate |
| CN109811040A (en) * | 2019-02-21 | 2019-05-28 | 北京农业智能装备技术研究中心 | A kind of germ quantitative detecting method based on PMA dyeing |
Non-Patent Citations (4)
| Title |
|---|
| TING CHEN等: "Transmission efficiency of Cotton leaf curl Multan virus by three cryptic species of Bemisia tabaci complex in cotton cultivars", 《PEERJ》 * |
| 王利华: "烟粉虱对高效氯氰菊酯和阿维菌素抗性的生化和分子机理", 《中国博士学位论文全文数据库（电子期刊）》 * |
| 章松柏等: "福州市发生由木尔坦棉花曲叶病毒引起的朱槿曲叶病", 《植物保护》 * |
| 陈婷等: "本地烟粉虱隐种加速入侵棉花曲叶病毒的广泛传播与危害", 《第五届全国入侵生物学大会—入侵生物与生态安全》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112048573A (en) * | 2020-09-28 | 2020-12-08 | 广东省农业科学院植物保护研究所 | RPA primer and kit for detecting cotton leaf curl virus, detection method and application thereof |
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