CN103563897A - Method for reducing plant virus carrying rate in insect medium by utilizing gossypol - Google Patents
Method for reducing plant virus carrying rate in insect medium by utilizing gossypol Download PDFInfo
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- CN103563897A CN103563897A CN201310291677.0A CN201310291677A CN103563897A CN 103563897 A CN103563897 A CN 103563897A CN 201310291677 A CN201310291677 A CN 201310291677A CN 103563897 A CN103563897 A CN 103563897A
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- bemisia tabaci
- gossypol
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- leaf curl
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Abstract
The invention discloses a method for reducing tomato yellowing leaf curl virus carrying rate in bemisia tabaci by utilizing gossypol. Aiming at solving the problem that a method for reducing the tomato yellowing leaf curl virus carrying rate in the bemisia tabaci to improve a disease control effect does not exist, gossypol is utilized for treating the bemisia tabaci, and the tomato yellowing leaf curl virus carrying rate in insect pests can be obviously reduced, so that the chance that the bemisia tabaci transmits viruses is reduced, and enhancement of the disease control effect can be facilitated. Compared with the traditional tomato yellowing leaf curl disease prevention and control means that a disease-pest-resistant variety, a chemical or physical means are mainly utilized for controlling quantity of the bemisia tabaci and viruses are controlled by adopting a virus antagonist in production, the method for reducing the tomato yellowing leaf curl virus carrying rate in the bemisia tabaci has the advantages that the gossypol is utilized for reducing plant virus carrying rate in the bemisia tabaci, so that a virus control effect is improved, an insect pest control manner is novel, safety is high, and the method for reducing the tomato yellowing leaf curl virus carrying rate in the bemisia tabaci by utilizing the gossypol is expected to have a certain application prospect.
Description
One, technical field
The invention belongs to techniques of pest control field, be specifically related to a kind of method of utilizing gossypol to reduce tomato yellow leaf curl virus carrying rate in Bemisia tabaci body.
Two, technical background
Bemisia tabaci Bemisia tabaci is a kind of worldwide insect, and it stings on the one hand inhales crop and cause directly and cause harm, and propagates on the other hand plant virus, causes being very popular of virus disease, to agricultural production, brings crushing disaster.Tomato yellow leaf curl virus (Tomato Yellow LeafCurl Virus, TYLCV) be the louse-borne most important plant virus of tobacco powder, it is strand ring-type DNA virus ，Shu geminivirus infection section (Geminiviridae) Begomovirus (Begomovirus) virus.Because causing it, the invading property of trunk offering of Bemisia tabaci generally occurring all over the world, tomato yellow leaf curl virus disease is also therefore popular in regional a plurality of countries such as Africa, America, Asia, most of provinces and cities such as Jiangsu, Zhejiang, Yunnan in China, have been spread to, not only make tomato drop in production over a large area, and its quality is declined.Except harm tomato, TYLCV also infects other plants such as capsicum, tobacco and pumpkin.
Once Bemisia tabaci is obtained after TYLCV, can retain all the life in vivo.If effectively reduce retaining of the interior TYLCV of Bemisia tabaci body, can significantly improve the control effect of virus disease.Yet in current production, the prevention and control of tomato yellow leaf curl are mainly taked to utilize Resistant kind, by chemistry or physical means, controlled Bemisia tabaci quantity, with viral antagonist, controlled the means such as virus, there is not yet and how effectively reduce Bemisia tabaci body inner virus carrying rate to improve the research report of Controlling effect.
Gossypol (gossypol) is a kind of polyatomic phenol material extracting in Malvaceae plant cotton, tree cotton or upland cotton mature seed, root skin, is peculiar secondary substance in cotton body.Found that gossypol is relevant with bt-cotton characteristic, also found that gossypol medically has anticancer, contraception and the effect such as antiviral.Whether does gossypol also have effect to plant virus? yet there are no research report both at home and abroad.The impact of research gossypol on tomato yellow leaf curl virus in Bemisia tabaci body, usings and expects to utilize gossypol as a kind of plant virus antagonist, aspect viroses of plant prevention and control, is playing a role.
Three, summary of the invention
1, goal of the invention
A kind of TYLCV method of removaling in desirable Bemisia tabaci body is provided, can reduces safely the carrying rate of the interior TYLCV of Bemisia tabaci body, for the control of tomato yellow leaf curl provides new approaches.
2, technical scheme
The present invention is a kind of removal method of insect amboceptor implants virus, it is characterized in that utilizing gossypol to reduce the carrying rate of TYLCV in Bemisia tabaci body.
3, beneficial effect
Method of the present invention compared with prior art, produce following beneficial effect: (1) is compared to control tomato yellow leaf curl with conventional control Bemisia tabaci quantity, the present invention controls virus disease by effective reduction Bemisia tabaci body implants virus carrying rate, and prevention and control thinking is new and with strong points; (2) compare with Agro-chemicals control Bemisia tabaci or virus disease, the present invention utilizes gossypol to control, and has conscientiously improved the nuisanceless level of control that virus disease is controlled, and effectively reduces environmental pollution.
Four, embodiment
Embodiments of the invention are the methods that adopt indoor measurement.
Assay method is as follows:
1, Bemisia tabaci connects poison
Gather Bemisia tabaci nymph at advanced age on same crop, after emergence, by the Bemisia tabaci of just sprouting wings, connect poison.Bemisia tabaci is placed on to tomato yellow leaf curl and falls ill on serious tomato plant and take food, with bagging, Bemisia tabaci be enclosed within to morbidity and without on worm blade, after the indoor 24h of taking food, take out.1 bag of every blade cover, 35 adults of every bag of cover, totally 16 bags.Wherein 4 bags of examination worms are positioned in-20 ℃ of refrigerators after taking food 24h, are directly used in virus and detect, and all the other examination worms are for gossypol Processing Test below.
2, gossypol is processed Bemisia tabaci
Adopt nutrient solution feeding method to carry out gossypol processing to Bemisia tabaci.By gossypol (standard items, Sigma company product) be dissolved in and in acetone, be made into mother liquor, and then mix with nutrient solution, preparation is containing the nutrient solution of 500mg/L and two kinds of concentration gossypols of 62.5mg/L respectively, for feeding, raise Bemisia tabaci, simultaneously with not containing gossypol but containing with 500mg/L gossypol nutrient solution in the nutrient solution of same amount acetone compare, feed and raise 6 days altogether.The logical glass tube in application two is tested, and 100 μ L nutrient solutions is added in one end sealed membrane interlayer of the logical glass tube in two, and Bemisia tabaci is put into pipe, and the other end seals with gauze.30 of every pipes, every processing 4 pipes.
Every two days during this time Ensure Liquid liquid once, added at the 1st day, the 3rd day and the 5th day respectively, added 100 μ L at every turn.
3, the detection of TYLCV in Bemisia tabaci
By the feeding Bemisia tabaci taking-up of the interior survival of each pipe afterwards in 6 days, put into refrigerator-20 ℃, for TYLCV, detect.
Single head Bemisia tabaci DNA extracts: in the centrifuge tube of 1.5ml, add the DNA alkaline lysis liquid (50mmol/LTris-HCl (pH8.0), 20mmol/LNaCl, 1mmol/LEDTA, 1%SDS) of 20 μ L, with sterilizing toothpick, fully grind.Add 1 μ L Proteinase K (20mg/ml), centrifugal 30 seconds, 60 ℃ of water-baths 1 hour, appended 1 μ L Proteinase K, and centrifugal 30s continues water-bath 2 hours, adds the distilled water of 178 μ L, 100 ℃ of water-bath 5min, inactivated proteases K.Get 100 μ L extracts in another centrifuge tube, add the precooling absolute ethyl alcohol of 2 times of volumes, fully mix, 4 ℃ of refrigerators precipitation centrifugal 10 minutes of rear 4 ℃ of 13000rpm that spend the night, pour out ethanol, add the distilled water dissolving DNA of 20 μ L ,-20 ℃ of preservations after drying.
The Auele Specific Primer that detects TYLCV is TV_P833 (5 '-GGTCTACACGCTTACGCCTTATT-3 ') and PATY_R (5 '-TTCCATCCGAACATTCAGGCAGC-3 '), and amplified fragments is approximately about 800bp.The reaction system of PCR is 10 μ L, comprises 5 μ Lmix (100mM KCl, 20mM Tris-HCl, 3mM MgCl
2, 400 μ M dNTPs, bromophenol blue), 0.2 μ L taq enzyme (5U/ μ L), 0.5 μ L primer TV_P833,0.5 μ L primer PATY_R, 3.2 μ L H
2o, 0.6 μ LDNA template.PCR parameter is: 95 ℃ of denaturation 5min, and 94 ℃ of sex change 45s, 56 ℃ of annealing 45s, 72 ℃ are extended 1min, 35 circulations, circulation finishes latter 72 ℃ and extends 20min.PCR product carries out electrophoresis and cloning and sequencing, defines its viral species.A viral Bemisia tabaci number is carried in each processing of record, calculates viral carrying rate.
With DPS software, respectively process viral carrying rate statistical analysis comparison.
Implementation result:
Research finds that Bemisia tabaci can significantly reduce the carrying rate (table 1) of TYLCV in its body after gossypol is processed.Having compared firm emergence Bemisia tabaci adults obtains after poison through TYLCV morbidity seedling indoor, the variation of TYLCV carrying rate in body after gossypol is processed again, find that Bemisia tabaci adults connects after poison on morbidity tomato seedling, in its body, TYLCV carrying rate is 100%, again through not feeding after 6d containing gossypol nutrient solution, the carrying rate of its TYLCV is 96.9%, both differences are not remarkable, and take food containing after 62.5mg/L gossypol nutrient solution, in Bemisia tabaci body, TYLCV carrying rate is 44.8%, extremely significantly lower than taking food the carrying rate that does not contain gossypol nutrient solution processed group, take food containing after 500mg/L gossypol nutrient solution, in Bemisia tabaci body, TYLCV carrying rate is lower, be only 17.4%, Zhuo is extremely significantly lower than taking food containing 62.5mg/L gossypol nutrient solution and not containing the carrying rate (F=188.446 of gossypol nutrient solution processed group, df=3, 15, P < 0.0001).
Table 1 gossypol is processed TYLCV carrying rate in rear Bemisia tabaci body
Note: after data, the formed objects mother stock that writes does not represent that in 1% and 5% level, difference is not remarkable.
Claims (2)
1. the method that insect amboceptor implants virus carrying rate reduces, is characterized in that utilizing gossypol to reduce the carrying rate of tomato yellow leaf curl virus in Bemisia tabaci body.
2. the gossypol that utilizes according to claim 1 reduces Bemisia tabaci body inner virus carrying rate, it is characterized in that 62.5mg/L gossypol processing insect, can significantly reduce insect virus carrying rate.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107173384A (en) * | 2017-05-24 | 2017-09-19 | 江苏省农业科学院 | A kind of method that utilization gossypol acetate removes plant virus in tomato plant |
| CN107279172A (en) * | 2017-06-29 | 2017-10-24 | 山东大农药业有限公司 | A kind of granule containing AVM and imidacloprid |
| CN108990981A (en) * | 2018-09-11 | 2018-12-14 | 江苏省农业科学院 | A method of Bemisia tabaci, which is removed, using Paeonol passes plant virus |
| CN109105366A (en) * | 2018-09-11 | 2019-01-01 | 江苏省农业科学院 | A method of utilizing gossypol degradation Sweet Potato Feathery Mottle Virus capsid protein |
| CN109169652A (en) * | 2018-09-11 | 2019-01-11 | 江苏省农业科学院 | A method of tomato yellow leaf curl virus is removed using phloroglucin |
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| CN102928598A (en) * | 2012-10-30 | 2013-02-13 | 浙江大学 | Dot-ELISA (dot Enzyme-Linked Immunosorbent Assay) method and tissue printing ELISA method for detecting presence of tomato yellow leaf curl virus in plant as well as reagent kit and application thereof |
| US20130247451A1 (en) * | 2011-12-27 | 2013-09-26 | Commonwealth Scientific And Industrial Research Organisation | Processes for producing lipids |
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2013
- 2013-07-12 CN CN201310291677.0A patent/CN103563897B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20130247451A1 (en) * | 2011-12-27 | 2013-09-26 | Commonwealth Scientific And Industrial Research Organisation | Processes for producing lipids |
| CN102928598A (en) * | 2012-10-30 | 2013-02-13 | 浙江大学 | Dot-ELISA (dot Enzyme-Linked Immunosorbent Assay) method and tissue printing ELISA method for detecting presence of tomato yellow leaf curl virus in plant as well as reagent kit and application thereof |
Non-Patent Citations (1)
| Title |
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| MARISA SKALJAC ET AL: "Tomato yellow leaf curl disease and plant–virus vector interactions", 《ISRAEL JOURNAL OF PLANT SCIENCES》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107173384A (en) * | 2017-05-24 | 2017-09-19 | 江苏省农业科学院 | A kind of method that utilization gossypol acetate removes plant virus in tomato plant |
| CN107279172A (en) * | 2017-06-29 | 2017-10-24 | 山东大农药业有限公司 | A kind of granule containing AVM and imidacloprid |
| CN108990981A (en) * | 2018-09-11 | 2018-12-14 | 江苏省农业科学院 | A method of Bemisia tabaci, which is removed, using Paeonol passes plant virus |
| CN109105366A (en) * | 2018-09-11 | 2019-01-01 | 江苏省农业科学院 | A method of utilizing gossypol degradation Sweet Potato Feathery Mottle Virus capsid protein |
| CN109169652A (en) * | 2018-09-11 | 2019-01-11 | 江苏省农业科学院 | A method of tomato yellow leaf curl virus is removed using phloroglucin |
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