CN101037673A - Tobacco virus abrasive paper frictional inoculation method - Google Patents
Tobacco virus abrasive paper frictional inoculation method Download PDFInfo
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- CN101037673A CN101037673A CN 200710077661 CN200710077661A CN101037673A CN 101037673 A CN101037673 A CN 101037673A CN 200710077661 CN200710077661 CN 200710077661 CN 200710077661 A CN200710077661 A CN 200710077661A CN 101037673 A CN101037673 A CN 101037673A
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- inoculation
- tobacco
- virus
- abrasive paper
- disease
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- 238000011081 inoculation Methods 0.000 title claims abstract description 65
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 46
- 241000700605 Viruses Species 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 39
- 244000061176 Nicotiana tabacum Species 0.000 title 1
- 241000208125 Nicotiana Species 0.000 claims abstract description 46
- 201000010099 disease Diseases 0.000 claims abstract description 33
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 33
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000012153 distilled water Substances 0.000 claims abstract description 6
- 239000004570 mortar (masonry) Substances 0.000 claims abstract description 5
- 208000024891 symptom Diseases 0.000 claims abstract description 4
- 241000196324 Embryophyta Species 0.000 claims description 19
- 244000137852 Petrea volubilis Species 0.000 claims description 16
- 241000723873 Tobacco mosaic virus Species 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 9
- 238000012360 testing method Methods 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 7
- 244000061456 Solanum tuberosum Species 0.000 claims description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 6
- 240000008067 Cucumis sativus Species 0.000 claims description 5
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims description 5
- 230000003612 virological effect Effects 0.000 claims description 5
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 210000003462 vein Anatomy 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 241000894007 species Species 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 8
- 208000034656 Contusions Diseases 0.000 abstract 1
- 238000007598 dipping method Methods 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 abstract 1
- 238000001914 filtration Methods 0.000 abstract 1
- 230000002265 prevention Effects 0.000 abstract 1
- 241000723762 Potato virus Y Species 0.000 description 7
- 208000015181 infectious disease Diseases 0.000 description 7
- 239000003082 abrasive agent Substances 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000002458 infectious effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- HBMJWWWQQXIZIP-UHFFFAOYSA-N silicon carbide Chemical compound [Si+]#[C-] HBMJWWWQQXIZIP-UHFFFAOYSA-N 0.000 description 3
- 229910010271 silicon carbide Inorganic materials 0.000 description 3
- 239000005723 virus inoculator Substances 0.000 description 3
- 241000724252 Cucumber mosaic virus Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 230000007115 recruitment Effects 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000011095 buffer preparation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229910001651 emery Inorganic materials 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
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- Cigarettes, Filters, And Manufacturing Of Filters (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a abrasive paper rubbing method of tobacco virus disease, including steps: (1) bruising leaf tissue with typical symptom of tobacco virus disease in mortar, adding water, filtering by gauze and extruding juice, then constant volume by distilled water to obtain inoculation liquor, compouded when need; (2) dividing a abrasive paper into equal portions and folding as arris; (3) dipping arris of the abrasive paper with inoculation liquor, sliding from one end to the other end of the arris on inoculation leaf, finishing inoculation. The method gets material easily, has simple, quick operation with little inaccuracy, only has three steps, success ratio of inoculation is 100%. The method suits for tobacco virus disease infected by machinery juice such as TMV, CMV, PVY, PVX and TEV and so on, especially for selecting and identifying fastness of tobacco species and experiment of prevention and cure tobacco virus disease.
Description
Technical field
The present invention relates to the inoculation method of tobacco diseases, inoculation method is propagated in the mechanical friction that is specifically related to tobacco virus.
Technical background
Inoculation test is the research of plant disease most important work, and, disease control popular to pathogen resistance, Disease Cycle, disease the research plant, infection processs and the pathogenic all respects inoculation tests such as differentiation of pathogen are prerequisites.Tobacco mosaic virus (tmv) (Tobacco Mosaic Virus, TMV), tobacco cucumber mosaic disease (Cucumber Mosaic Virus, CMV) and tobacco potato Y virus disease (Potato Virus Y, PVY) be tobacco main diseases viral disease, these tobacco virus have the mechanical friction route of infection, the infectious tobacco virus inoculation method of mechanical friction mainly contains diseased plant juice frictional inoculation method now, the spray gun inoculation method, diseased tissues direct inoculation etc., with regard to present tobacco virus inoculation method, under the very many situations of inoculation material, the problem of every kind of inoculation method just comes out, and is big as inoculation virus quantity error between plant, waste time and energy and operation easier bigger etc.Diseased plant juice frictional inoculation method is the most frequently used inoculation method, and its step is to get disease leaf texture to grind in mortar, adds a spot of water, and is with filtered through gauze and extrude juice, standby with the certain ratio of dilute phosphoric acid buffered soln dilution at last.During inoculation, be added in a small amount of fine emery powder in the juice or be sprinkling upon on the blade face of inoculation as abrasive material, use finger, gauze, writing brush or little flat brush etc. then, dip in taking juice friction gently back and forth on blade, again with clear water juice that the blade face is unnecessary and abrasive material flush away gently.Its shortcoming is that program is loaded down with trivial details, wastes time and energy, and inoculation virus quantity error is big between plant, and is particularly particularly evident in more than big area inoculation and the material.The spray gun inoculation method is to add silicon carbide in diseased plant juice, in the identical time, with identical spraying pressure, the diseased plant juice that will contain a small amount of silicon carbide is beaten and is being inoculated by on the plant that inoculates, its shortcoming is that operation easier is bigger, is controlled at identical time and uniform pressure and lacks certain operability; Lance driving device is expensive, makes experimentation cost higher.Diseased tissues is directly inoculated on the blade face, claims again " dry receive ", be applicable to a small amount of inoculation or diseased tissues material seldom with the unsettled situation of virus, its shortcoming be can't carry out a large amount of inoculation tests, infectious rate is lower and plant between inoculation virus quantity error big.
Through retrieval, the method that does not also adopt sand paper to inoculate is come out, and does not yet have new inoculation method to apply for a patent.
Summary of the invention
The object of the present invention is to provide a kind of tobacco virus abrasive paper frictional inoculation method, enable to reduce inoculation virus quantity between plant error, reduce experimental cost, realizes simple to operate, quick, the raising infectious rate, thus overcome the defective of prior art.
The contriver is engaged in tobacco mosaic virus (tmv) (Tobacco Mosaic Virus for a long time, TMV), tobacco cucumber mosaic disease (Cucumber Mosaic Virus, CMV) and tobacco potato Y virus disease (Potato Virus Y, PVY) in the research of tobacco bred resistance screening and aspects such as evaluation and control thereof, these tobacco virus (as TMV, CMV, PVY and TEV etc.) have the juice mechanical friction route of infection.The contriver is combined in the reality of grass-roots unit and inoculation lot of materials, is improved to tobacco virus abrasive paper frictional inoculation method based on the defective of above-mentioned inoculation method.
The tobacco virus abrasive paper frictional inoculation method that the contriver provided may further comprise the steps:
1. get the blade of typical tobacco virus disease symptoms, in mortar, grind, add water, with filtered through gauze and extrude juice, use the distilled water constant volume then, obtain inoculation liquid, now with the current;
2. get commercially available sand paper, directly use or morsel, be folded to form rib;
3. dip in the rib of sand paper and get inoculation liquid and on the inoculation plant leaf, draw the other end from an end of rib gently, promptly finish inoculation.
In the first step of above-mentioned seeded process, the amount that adds water arrives to be convenient to tissue grinding and to get final product; Described constant volume is to weigh behind the vein of picking out the disease leaf, press then mass volume ratio (W: V) with distilled water diluting to certain proportion, it is as follows that typical virus disease leaf tissue grinds the juice Dilution ratio:
Tobacco mosaic virus (tmv) (TMV) 1: 50～100 (W/V)
Tobacco potato Y virus disease (PVY) 1: 10～50 (W/V)
Tobacco cucumber mosaic disease (CMV) 1: 5～10 (W/V)
In second step of above-mentioned seeded process, usually commercially available sand paper is divided into the fritter of 4 homalographics, be folded to form rib, block-by-block uses.
In the 3rd step of above-mentioned seeded process, dip in the sand paper rib and to get number of times that inoculation liquid rows dry and decide, but that each inoculates the number of times that plant draws is identical according to concrete inoculation test and viral species.
The contriver points out: the sand paper material is easy to get, cost is low, and the sand face is equivalent to silicon carbide, plays the effect of abrasive material, can increase virus and invade wound and p of E, improves infection rate and rate of vaccination; Because the sand paper area equates that the plant that has only the rib contact to inoculate can guarantee that like this virus inoculation amount is in the limit of error of minimum; And now with the current inoculation liquid is obvious to unsettled viral effect, also can reduce the buffer preparation step.
Adopt tobacco virus abrasive paper frictional inoculation method of the present invention, draw materials easily, simple to operate, quick, error is little, operation steps had only for three steps, and success ratio of inoculation is 100%.Be applicable to the virus disease that tobacco mosaic virus (tmv) (TMV), tobacco cucumber mosaic disease (CMV), tobacco potato Y virus disease (PVY), tobacco potato X virus disease (PVX) and etch virus of tobacco viral disease (TEV) etc. are propagated by mechanical friction.It is very obvious that present method is carried out the virus disease field test effect in grass-roots unit, is specially adapted to the test of tobacco bred resistance screening and evaluation and tobacco virus thereof control medicament.
Description of drawings
Fig. 1 is the sand paper floor map, and Fig. 2 is the folding back of sand paper synoptic diagram.1 is the sand face among the figure, and 2 is the sand paper diagonal lines, and 3 is paper, and 4 is rib.
Embodiment
Embodiment 2004～2006 years carries out the sick field control effectiveness test of tobacco mosaic virus (tmv) every year in the Dafang County, Guizhou Province, the flue-cured tobacco on ground has 3500 strain flue-cured tobaccos approximately more than 3 mu; 2004～2006 years, all carried out mosaic disease screening varieties and evaluation in Jinsha County, Guizhou Province Chinese tobacco breeding breeding base every year, and test materials has about 5 mu of ground, about 5000～6000 strain flue-cured tobaccos.
1. get blade (the picking out vein) 20g of typical tobacco mosaic virus (tmv) symptom, add less water and in mortar, grind, with filtered through gauze and extrude juice, use 1～5L distilled water constant volume then, obtain inoculation liquid;
2. get 2 on commercially available sand paper, be divided into 8 fritters of homalographic, be folded to form rib;
3. dip in the rib of sand paper and get inoculation liquid and on the inoculation plant leaf, draw the other end from an end of rib gently, promptly finish inoculation.
Adopt tobacco virus abrasive paper frictional inoculation method, a people is inoculated into TMV on the 1000 strain flue-cured tobaccos, finishes nearly about 1 hour of the operating time of inoculation; And, inoculate 1000 strain flue-cured tobaccos with original diseased plant juice rubbing manipulation inoculation, 4 people's operations wanted finish in nearly about 1 hour.Tobacco virus abrasive paper frictional inoculation method and diseased plant juice frictional inoculation method have been saved 3 times of recruitments on recruitment efficient; The tobacco virus abrasive paper inoculation method does not have technical requirements substantially, and is simple to operate, quick, and interpersonal inoculation error is not remarkable yet, and success ratio of inoculation is 100%.
Claims (4)
1 tobacco virus abrasive paper frictional inoculation method is characterized in that this method may further comprise the steps:
The sick leaf texture of 1. getting typical mosaic disease symptom grinds in mortar, adds water, with filtered through gauze and extrude juice, uses the distilled water constant volume then, obtains inoculation liquid, and is now with the current;
2. get commercially available sand paper, directly use or morsel, be folded to form rib;
3. dip in the rib of sand paper and get inoculation liquid and on the inoculation plant leaf, draw the other end from an end of rib gently, promptly finish inoculation.
2 according to the described tobacco virus abrasive paper frictional inoculation method of claim 1, it is characterized in that in the first step of above-mentioned seeded process, and amount of water arrival is convenient to the tissue grinding and is got final product; Described constant volume is to weigh behind the vein of picking out the disease leaf, press then mass volume ratio with distilled water diluting to certain proportion, it is as follows that typical virus disease leaf tissue grinds the juice Dilution ratio:
Tobacco mosaic virus (tmv) 1: 50～100
Tobacco potato Y virus disease 1: 10～50
Tobacco cucumber mosaic disease 1: 5～10.
3 according to the described tobacco virus abrasive paper frictional inoculation method of claim 1, it is characterized in that in second step of above-mentioned seeded process commercially available sand paper being divided into the fritter of equal portions, is folded to form rib, and block-by-block uses.
4 according to the described tobacco virus abrasive paper frictional inoculation method of claim 1, it is characterized in that in the 3rd step of above-mentioned seeded process, dip in the rib of sand paper and to get number of times that inoculation liquid draws and decide, but that each inoculates the number of times that plant draws is identical according to concrete inoculation test and viral species.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100776614A CN101037673B (en) | 2007-02-15 | 2007-02-15 | Tobacco virus disease sand paper friction inoculation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100776614A CN101037673B (en) | 2007-02-15 | 2007-02-15 | Tobacco virus disease sand paper friction inoculation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101037673A true CN101037673A (en) | 2007-09-19 |
| CN101037673B CN101037673B (en) | 2012-04-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100776614A Expired - Fee Related CN101037673B (en) | 2007-02-15 | 2007-02-15 | Tobacco virus disease sand paper friction inoculation method |
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| Country | Link |
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| CN (1) | CN101037673B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972218A (en) * | 2012-11-16 | 2013-03-20 | 中国农业科学院烟草研究所 | Tobacco virus disease quick rubbing inoculation method |
| CN106508335A (en) * | 2016-09-23 | 2017-03-22 | 湖北省农业科学院经济作物研究所 | Identification method for black rot resistance of alpine radish in seedling stage |
| CN106636465A (en) * | 2016-12-30 | 2017-05-10 | 中国烟草总公司广东省公司 | Rapid PVY virus identification method and application thereof |
| CN106702023A (en) * | 2016-12-30 | 2017-05-24 | 中国烟草总公司广东省公司 | Rapid identification method of PVX viruses and application thereof |
| CN107988173A (en) * | 2017-11-24 | 2018-05-04 | 贵州省烟草科学研究院 | One grows tobacco CMV inoculation methods |
| CN111066516A (en) * | 2019-12-19 | 2020-04-28 | 扬州大学 | A kind of rapid inoculation method of seedling leaves of barley yellow mosaic virus |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100355739C (en) * | 2005-07-05 | 2007-12-19 | 南开大学 | Diazosulfide derivative, and its synthesizing method and activity for inducing tobacco to resist tobacco mosaic virus |
-
2007
- 2007-02-15 CN CN2007100776614A patent/CN101037673B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972218A (en) * | 2012-11-16 | 2013-03-20 | 中国农业科学院烟草研究所 | Tobacco virus disease quick rubbing inoculation method |
| CN106508335A (en) * | 2016-09-23 | 2017-03-22 | 湖北省农业科学院经济作物研究所 | Identification method for black rot resistance of alpine radish in seedling stage |
| CN106636465A (en) * | 2016-12-30 | 2017-05-10 | 中国烟草总公司广东省公司 | Rapid PVY virus identification method and application thereof |
| CN106702023A (en) * | 2016-12-30 | 2017-05-24 | 中国烟草总公司广东省公司 | Rapid identification method of PVX viruses and application thereof |
| CN107988173A (en) * | 2017-11-24 | 2018-05-04 | 贵州省烟草科学研究院 | One grows tobacco CMV inoculation methods |
| CN111066516A (en) * | 2019-12-19 | 2020-04-28 | 扬州大学 | A kind of rapid inoculation method of seedling leaves of barley yellow mosaic virus |
| CN111066516B (en) * | 2019-12-19 | 2022-04-22 | 扬州大学 | A kind of rapid inoculation method of seedling leaves of barley yellow mosaic virus |
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| Publication number | Publication date |
|---|---|
| CN101037673B (en) | 2012-04-11 |
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Granted publication date: 20120411 Termination date: 20160215 |