CN104673800A - Method for preventing and treating fruit tree RNA virus - Google Patents
Method for preventing and treating fruit tree RNA virus Download PDFInfo
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- CN104673800A CN104673800A CN201510126348.XA CN201510126348A CN104673800A CN 104673800 A CN104673800 A CN 104673800A CN 201510126348 A CN201510126348 A CN 201510126348A CN 104673800 A CN104673800 A CN 104673800A
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Abstract
The invention relates to a method for preventing and treating a fruit tree RNA virus and belongs to the technical field of biology. The method comprises the following steps: (1) designing the siRNAs of the ACLSV, ASPV and ASGV according to part of conserved sequences of the ACLSV, ASPV and ASGV coat protein genes; (2) preparing 0.1mu molL-1.2mu mol/L siRNA double distilled water mixed solution containing ACLSV, ASPV and ASGV, wherein the mass ratio of the ACLSV, ASPV to ASGV is 1:1:1; (3) adding 1-10% of the spreader tween-80 into the siRNA double distilled water mixed solution of the ACLSV, ASPV and ASGV; and (4) spraying the mixed solution obtained from the step (3) on the leaf surface of the fruit tree carefully and considerately. The siRNAs of the ACLSV, ASPV and ASGV are first synthesized and introduced into the apple tree and pear tree body through a leaf surface spraying method. The method has the advantages of simple operation method, rapid speed, strong instantaneity, obvious effect, low cost and easy popularization.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of method of preventing and treating RNA Viruses in Fruit Trees.
Background technology
Fruit diseases is fungal disease mainly, Micobial Disease and virus disease (comprising virus, viroid, rickettsia-like organism, bacterioid etc.) three major types.Current people are also not enough to virus disease understanding, pay attention to not.Along with the development of global fruit growing, fruit tree area constantly expands, fruit tree virus disease to adopt for a long time the fruit tree hazard area of nourishing and generating and degree also increasing, cause many fruit tree growths bad, production declining, product qualitative change is bad, and tree body surface reveals many abnormalities, bring huge financial loss to production of fruit trees, and this loss still has the trend constantly spread.
Apple chlorotic leaf spot virus (Apple chlorotic leaf spotvirus, ACLSV), apple stem grooving virus (Apple stemgrooving virus, and apple stem pitting virus (Apple stem pitting virus ASGV), ASPV) be the three large main viruses endangering apple tree and pear tree, three is RNA viruses.ACLSV, ASPV and ASGV present not reveal any symptoms on host, i.e. non-evident sympton in appearance, therefore are also called latent infection, are not easily found by people, only on highstrung plant indicator viral to this, just can show peculiar symptom.ACLSV, ASGV and ASPV are almost present in all apples and pears producing region, the latent band rate of some apple and Pear varieties is saturated, perplex the 1950's the Fuji apple of Japanese fruit tree circle high connect disease be proved to be by ACLSV, ASPV and ASGV separately or Combined Infection cause.In some fruit producing regions of China, also find that ACLSV, ASPV and ASGV are everlasting on certain Stockscion combination the eighties in 20 century and cause Incompatibility, in addition, shoot growth also can be caused abnormal, branch is sparse, tree vigo(u)r fails, and cause Severe Reduction, fruit quality is subject to baneful influence.
The virus disease of apple tree and pear tree belongs to refractory and treats disease, people also do not develop a kind of effectively preventing medicament for many years, comparatively feasible measure of control cultivate virus-free seedling and Non-toxic cultivation, nontoxic seedling cultivates the normal Micro-stem tip tissue culture that adopts in conjunction with heat treatment technics, coordinate sensitive simultaneously, efficiently, single-minded detection method, Non-toxic cultivation then needs the basis of cultivating at nontoxic seedling to be aided with strict field and selects, pruning, rich water quality managements etc. are a series of loaded down with trivial details, complicated cultivation condition, huge difficulty is brought to the preventing and controlling of apple and pears virus disease, not only preventing and controlling amount is large, complicated loaded down with trivial details, and cost accounting is high.
RNA perturbation technique (RNAi) is the FA Protocols in Molecular Biology of one grown up in recent years, is widely used in the aspects such as large-scale functional genome resolves, the disease-resistant and regulation and control of growing.As a kind of means realizing RNA interference, siRNA has the effect bringing out RNA interference in vivo, reaches the effect suppressing virus, prevent and treat virus disease thus.The application of current RNA perturbation technique in nematode, fruit bat and plant has achieved a lot of achievement, and the investigation and application in Mammals also obtains incremental advances.The gene silencing of RNA mediation is the self-protective mechanism of plant in gene regulating level, plays a significant role, have ubiquity in antagonism virus infection, DNA intrusion, DNA swivel base and rearrangement.
Summary of the invention
The present invention, for solving the problems such as fruit tree virus disease prevention and controls in prior art is loaded down with trivial details, complicated, provides a kind of based on modern biotechnology, the method for control RNA Viruses in Fruit Trees easy and simple to handle, speed is fast, cost is low, rapid-action, real-time.
Inventive point of the present invention is: control RNAi being introduced apple and pears virus disease, according to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene.After utilizing plant stomata and epidermic cell absorption siRNA, siRNA to enter cell, degraded the mRNA of virus coat protein gene by RNA interference principle, virus cannot be copied, serve the effect suppressing virus, prevent and treat virus disease.
First object of the present invention is a kind of siRNA sequence preventing and treating RNA Viruses in Fruit Trees of protection, and the siRNA sequence of ACLSV is as shown in SEQ ID NO.1, and the siRNA sequence of ASGV is as shown in SEQ ID NO.2, and the siRNA sequence of ASPV is as shown in SEQ ID NO.3.
Another object of the present invention is a kind of method of preventing and treating RNA Viruses in Fruit Trees of protection, comprises the following steps:
(1) according to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ',
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ',
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
(2) compound concentration is the siRNA distilled water mixing solutions containing ACLSV, ASPV and ASGV of 0.1 μm of ol/L ~ 1.2 μm ol/L; Wherein the quality proportioning of the siRNA of ACLSV, ASPV and ASGV is 1:1:1;
(3) in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, add spreader-sticker tween-80, wherein the percent by volume of spreader-sticker tween-80 is 1% ~ 10%;
(4) step (3) gained mixed solution is sprayed the blade surface in fruit tree, solution spraying is careful, thoughtful.
Further, described step (4) is in annual fruit tree growth in 6 ~ August vigorous period, and the blade surface to fruit tree sprays step (3) gained mixed solution.
Further, described fruit tree is apple tree or pear tree.
The present invention adopts the RT-PCR system set up the apple tree and pear tree that sprayed siRNA mixing solutions to be carried out to the detection of ACLSV, ASPV and ASGV, detected result shows all to carry ACLSV, ASPV and ASGV in the tree body before testing, and the result detected in different time sampling after experiment all shows not carry ACLSV, ASPV and ASGV in tree body.Prove thus, the present invention has achieved good prevention effect, the apple easy as a kind of working method, successful, use cost are low and pears virus disease control method, adopts RNAi control apple and pears virus disease to have a good application prospect.
Compared with prior art, beneficial effect is in the present invention: (1) adopts RNA perturbation technique to prevent and treat ACLSV, ASPV and ASGV virus disease first; (2) siRNA of ACLSV, ASPV and ASGV is synthesized first; (3) adopt foliage-spray method, with the tween-80 (Tween-80) of percent by volume 1 ~ 10% as spreader-sticker, the siRNA of ACLSV, ASPV and ASGV is introduced in apple tree and pear tree body; (4) working method of the present invention is easy, and speed is fast, real-time, successful, and use cost is low, is easy to promote.
Embodiment
Below in conjunction with embodiment, the invention will be further described.Following examples are only for illustration of the present invention but not for limiting scope of the present invention, in embodiment, the experimental technique of unreceipted actual conditions, conventionally operates usually.
In embodiment 1 ~ 6, the apple tree kind that each embodiment is selected has: Fuji apple, Jin Guan, new Red Star, Qiao Najin and Qin Guan totally 5 kinds, and each Fruit variety 5 strain tree, amounts to 150 strains; The Pear Trees with Various Cultivars that each embodiment is selected has: three season pears, bar pears and pear totally 3 kinds, each Fruit variety 5 strain tree, amounts to 90 strains.
On pretreatment, win blade from every strain fruit tree some, be placed in curling stone cryopreservation, be convenient to test the later stage and carry out RT-PCR detection analysis.ACLSV, ASPV and ASGV Coat protein gene sequence is utilized to design primer, carry out reverse transcription (RT) and polymerase chain reaction (PCR) of total serum IgE, check siRNA to the prevention effect of ACLSV, ASPV and ASGV by above-mentioned RT-PCR detected result.
Embodiment 1
1, fruit tree is sprayed
Experimental period: on June 16th, 2009 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 0.1 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Some respectively on July 10th, 2010, on July 16th, 2012 and on July 25th, 2014 getting blade on experimental plants, be placed in curling stone cryopreservation, take back use for laboratory and detect analysis in RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 0.1 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Embodiment 2
1, fruit tree is sprayed
Experimental period: on July 20th, 2009 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 0.3 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Respectively on July 10th, 2010, on July 16th, 2012 and on July 25th, 2014 getting blade on experimental plants some, be placed in curling stone cryopreservation, take back laboratory and detect analysis for carrying out RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 0.3 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Embodiment 3
1, fruit tree is sprayed
Experimental period: on August 9th, 2009 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 0.5 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Respectively on July 10th, 2010, on July 16th, 2012 and on July 25th, 2014 getting blade on experimental plants some, be placed in curling stone cryopreservation, take back laboratory and detect analysis for carrying out RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 0.5 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Embodiment 4
1, fruit tree is sprayed
Experimental period: on June 20th, 2010 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 0.7 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Respectively on July 15th, 2011, on July 20th, 2012 and on July 18th, 2014 getting blade on experimental plants some, be placed in curling stone cryopreservation, take back laboratory and detect analysis for carrying out RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 0.7 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Embodiment 5
1, fruit tree is sprayed
Experimental period: on July 25th, 2010 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 0.9 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Respectively on July 15th, 2011, on July 20th, 2012 and on July 18th, 2014 getting blade on experimental plants some, be placed in curling stone cryopreservation, take back laboratory and detect analysis for carrying out RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 0.9 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Embodiment 6
1, fruit tree is sprayed
Experimental period: on August 13rd, 2010 is fine, at dusk 5 points;
Experiment place: the 12 years green apple arboretums in Jinzhou district of Dalian and certain 15 years raw pear tree gardens;
Each apple variety and Pear varieties respectively select 5 strain trees, and before experiment process, it is some that every strain tree wins blade, is placed in curling stone cryopreservation, analyze for carrying out RT-PCR detection.According to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ';
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ';
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
By the siRNA of ACLSV, ASPV and ASGV according to quality proportioning 1:1:1, be mixed with the distilled water mixing solutions of concentration 1.2 μm of ol/L; The spreader-sticker tween-80 that percent by volume is respectively 1%, 3%, 5%, 7% and 10% is added in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, be sprayed on respectively by the solution prepared on the blade surface of every strain tree, solution spraying is careful, thoughtful.
2, Viral diagnosis
Respectively on July 15th, 2011, on July 20th, 2012 and on July 18th, 2014 getting blade on experimental plants some, be placed in curling stone cryopreservation, take back laboratory and detect analysis for carrying out RT-PCR.Take 100 ~ 200mg to before every strain tree above spray respectively with the rear blade plucked of spray, repeat 3 times and weigh, the RT-PCR detection system that employing has been set up carries out the detection of ACLSV, ASPV and ASGV.Detected result shows: all carry ACLSV, ASPV and ASGV virus in the tree body before the siRNA mixing solutions of spraying 1.2 μm of ol/LACLSV, ASPV and ASGV, and the result detected in different time sampling after spraying all shows not carried ACLSV, ASPV and ASGV virus in all experimental plants tree body.
Table 1 embodiment 1 ~ 6 sprays Experimental comparison
Note: feminine gender is-, the positive is+.
As shown in table 1 is that embodiment 1 ~ 6 sprays experimental result contrast, before spraying, tree body all carries virus, after spraying, tree body all can't detect virus, prove that method of the present invention effectively can prevent RNA Viruses in Fruit Trees, and, this detection also finds, the siRNA mixed solution of ACLSV, ASPV and ASGV its result for the treatment of of virus disease is not limited by concentration, the siRNA mixed solution of ACLSV, ASPV and ASGV of different concns all effectively can prevent RNA Viruses in Fruit Trees.
Claims (4)
1. prevent and treat a siRNA sequence for RNA Viruses in Fruit Trees, it is characterized in that, the siRNA sequence of ACLSV is as shown in SEQ ID NO.1, and the siRNA sequence of ASGV is as shown in SEQ ID NO.2, and the siRNA sequence of ASPV is as shown in SEQ ID NO.3.
2. prevent and treat a method for RNA Viruses in Fruit Trees, it is characterized in that, comprise the following steps:
(1) according to the siRNA of the design of part conserved sequence ACLSV, ASPV and ASGV of ACLSV, ASPV and ASGV coat protein gene;
The siRNA sequence of described ACLSV is: 5 '-UUGGAGUUCAAUAAGGGUCUG-3 ',
The siRNA sequence of ASGV is: 5 '-AACCUGGGUCGCGGUCUUGGAC-3 ',
The siRNA sequence of ASPV is: 5 '-ACUGAGCGGUGUACGUUGAGGCA-3 ';
(2) compound concentration is the siRNA distilled water mixing solutions containing ACLSV, ASPV and ASGV of 0.1 μm of ol/L ~ 1.2 μm ol/L; Wherein the quality proportioning of the siRNA of ACLSV, ASPV and ASGV is 1:1:1;
(3) in the siRNA distilled water mixing solutions of ACLSV, ASPV and ASGV, add spreader-sticker tween-80, wherein the percent by volume of spreader-sticker tween-80 is 1% ~ 10%;
(4) step (3) gained mixed solution is sprayed the blade surface in fruit tree, solution spraying is careful, thoughtful.
3. a kind of method of preventing and treating RNA Viruses in Fruit Trees according to claim 2, is characterized in that, described step (4) is in annual fruit tree growth in 6 ~ August vigorous period, and the blade surface to fruit tree sprays step (3) gained mixed solution.
4. a kind of method of preventing and treating RNA Viruses in Fruit Trees according to claim 2, is characterized in that, described fruit tree is apple tree or pear tree.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105463014A (en) * | 2015-12-30 | 2016-04-06 | 中国农业科学院郑州果树研究所 | RNAi anti-ASPV (Apple Stem Pitting Virus) expression vector and application thereof in apple breeding for disease resistance |
| CN107372609A (en) * | 2017-07-27 | 2017-11-24 | 河南柏裕植物免疫科技有限公司 | Apple virus disease vaccine |
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| CN101914540A (en) * | 2010-08-09 | 2010-12-15 | 大连大学 | Method for introducing RNA interference into plants |
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| CN101914540A (en) * | 2010-08-09 | 2010-12-15 | 大连大学 | Method for introducing RNA interference into plants |
Non-Patent Citations (2)
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| 刘辉: "苹果茎沟病毒cp基因hpRNA表达载体的构建及其转化烟草研究", 《中国优秀硕士论文全文数据库 农业科技辑》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105463014A (en) * | 2015-12-30 | 2016-04-06 | 中国农业科学院郑州果树研究所 | RNAi anti-ASPV (Apple Stem Pitting Virus) expression vector and application thereof in apple breeding for disease resistance |
| CN105463014B (en) * | 2015-12-30 | 2019-01-01 | 中国农业科学院郑州果树研究所 | The anti-apple stem pitting virus expression vector of RNAi and its application in apple breeding for disease resistance |
| CN107372609A (en) * | 2017-07-27 | 2017-11-24 | 河南柏裕植物免疫科技有限公司 | Apple virus disease vaccine |
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