CN107360906A - A kind of method for mitigating sweet potato SPVD harm - Google Patents
A kind of method for mitigating sweet potato SPVD harm Download PDFInfo
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- CN107360906A CN107360906A CN201710662129.2A CN201710662129A CN107360906A CN 107360906 A CN107360906 A CN 107360906A CN 201710662129 A CN201710662129 A CN 201710662129A CN 107360906 A CN107360906 A CN 107360906A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
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Abstract
The present invention discloses a kind of method for mitigating sweet potato SPVD harm, belong to plant pest prevention and control field, use concentration for 0.1% Physcion, it is made into 300 times of dilutions, dispenser is carried out by the way of to sweet potato foliar spray, using the dilution of Physcion, Sweet Potato Leaf is sprayed weekly after sweet potato growing 1 time, continuously spray 4 weeks.The present invention sprays two kinds of viral levels of FMV and CSV in above-mentioned medicament rear blade by fluorescence quantifying PCR method measure, sweet potato physical signs and economical character is determined, to evaluate mitigation effect of the medicament to sweet potato SPVD.The present invention is sprayed on sweet potato blade face using 0.1% Physcion, can not only reduce production loss, moreover it is possible to significantly reduce relative amount viral in sweet potato, have preferable prevention effect to sweet potato SPVD.
Description
Technical field
The present invention relates to a kind of method for mitigating sweet potato SPVD harm, belong to plant pest prevention and control field.
Background technology
Sweet Potato Feathery Mottle Virus (sweet potato feathery mottle virus, SPFMV) and sweet potato chlorisis
Stunt virus (sweetpotato chlorotic stuntvirus, SPCSV) while infect sweet potato and during interaction, can produce sweet
Potato virosis complex (SPVD), has a strong impact on the production of sweet potato, is that maximum disease is endangered Sweet Potato Industry in world wide
One of.Sweet potato infection SPVD after, can produce blade face distortion, deformity, chlorisis, bright arteries and veins and plant dwarfing etc. symptom, sweet potato infect
Production loss is up to 80% after SPVD.
Sweet potato SPVD preventing and treatings at present mainly by Sweetpotato Viruses Elimination, kill communication media and spray antiviral agricultural chemicals etc..Pass through
The drawbacks such as cycle long, easy recurrence, cost height be present in poison-removing method control SPVD；Chemical agent such as alkene hydroxyl moroxydine, phonetic peptide
The medicaments such as mycin can suppress the synthesis of viral nucleic acid and lipoprotein, but the antiviral pesticide variety of in the market is various, to sweet potato SPVD
Effect selectivity and inhibition there is no system research.
The content of the invention
In view of the above-mentioned problems of the prior art, the present invention provides a kind of method for mitigating sweet potato SPVD harm, screening
Go out to significantly inhibit sweet potato SPVD medicament, the development to Sweet Potato Industry is significant.Invention additionally provides one kind
The evaluation method of the SPVD extents of injury.
To achieve these goals, the present invention use a kind of mitigation sweet potato SPVD harm method, use concentration for
0.1% Physcion, 300 times of dilutions are made into, dispenser is carried out by the way of to sweet potato foliar spray.
As an improvement, using the dilution of Physcion, Sweet Potato Leaf is sprayed weekly after sweet potato growing 1 time, continuously
Spray 4 weeks.
In addition, present invention also offers a kind of evaluation method of the sweet potato SPVD extents of injury, pass through fluorescence quantifying PCR method
Measure sprays two kinds of viral levels of FMV and CSV in above-mentioned medicament rear blade, sweet potato physical signs and economical character is determined, to comment
Mitigation effect of the valency medicament to sweet potato SPVD.
The present invention is sprayed on sweet potato blade face using 0.1% Physcion, can not only reduce production loss, moreover it is possible to significantly drop
Viral relative amount in low sweet potato, there is preferable prevention effect to sweet potato SPVD.
Brief description of the drawings
The RT-PCR detections that Fig. 1 is FMV and CSV in Sweet Potato Leaf；
Fig. 2 is the change for spraying FMV and CSV virus relative amounts in Sweet Potato Leaf after medicament；
Fig. 3 is the influence for spraying medicament to Sweet Potato Leaf Determination of Chlorophyll content；
Fig. 4 is the influence for spraying medicament to the climing length of sweet potato；
Fig. 5 is the influence for spraying medicament to sweet potato underground part fresh weight；
Fig. 6 is the influence for spraying medicament to sweet potato overground part fresh weight；
Fig. 7 is the influence for spraying medicament to sweet potato total biomass.
Embodiment
To make the object, technical solutions and advantages of the present invention of greater clarity, below by drawings and Examples, to this
Invention is further elaborated.However, it should be understood that the specific embodiments described herein are merely illustrative of the present invention,
The scope being not intended to limit the invention.
Unless otherwise defined, all technical terms and scientific terminology used herein are led with belonging to the technology of the present invention
The implication that the technical staff in domain is generally understood that is identical, and used term is intended merely to retouch in the description of the invention herein
State the purpose of specific embodiment, it is not intended that in the limitation present invention.
Experimental method used is conventional method unless otherwise specified in following embodiments.In following embodiments
Material used, reagent etc., unless otherwise specified, are commercially obtained.
Sweet potato variety is sweet potato variety Xu purple potato 8 of Xuzhou Sweetpotato Center's seed selection.Test material is derived from river
Su Xuzhou sweet potatoes research center chlorisis is downgraded, the plant of leaf malformation.
Medicament provided by the present invention is the medicament for being used to prevent and treat the viroses of plant obtained by commercial sources.
Embodiment 1
1. Sweet Potato Leaf Total RNAs extraction and RT-PCR detections：
Sweet Potato Leaf total serum IgE is extracted with CHMC's ultrafast novel plant RNA extracts kits in ocean, is removed with supporting DNA enzymatic
Genomic DNA, obtained using Takara Reverse Transcriptase M-MLV (RNase H) kit reverse transcription
CDNA, detected using health to enter performing PCR 2 × Taq of century Master Mix, primer is by Shanghai Sheng Gong bioengineering Co., Ltd
Synthesis.
FMV primer sequences are：
FM5CP–CCGGGTCTRRTGAGARHACTGAATTTAAAGATGC(SEQ ID NO.1)
FMVCP3–GACTCTCGAGCCTATTGCACACCCCTCATTCC(SEQ ID NO.2)
Target gene length is 945bp；
CSV primer sequences are：
CSV70P1–GACGGKGGTACKATGAARGTCC(SEQ ID NO.3)
CSV70P2-GGCTCACAAACHGAYTTCATAAACAT(SEQ ID NO.4)
Target gene length 431bp.
PCR primer is detected with 0.1% agarose gel electrophoresis, has amplified grown with SPFMV and SPCSV target gene respectively
Spend consistent band.It is century quick Ago-Gel DNA QIAquick Gel Extraction Kits CW2301 recovery DNA fragmentation with health, serves Hai Sheng
Work bioengineering Co., Ltd is sequenced.Sequencing result is analyzed through BLAST, the one of FMV sequences in target gene FMV and GenBank
Cause property is respectively 97.73%；The uniformity of CSV sequences is respectively 99.11% in target gene CSV and GenBank.The above results
Illustrate, selected sweet potato plant infects two kinds of viruses of SPFMV and SPCSV, as shown in Figure 1.
2. Sweet Potato Leaf SPVD detections and relatively viral assay after chemicals treatment：
The infection SPVD sweet potato plant detected through qRT-PCR are chosen, are sprayed weekly once by corresponding spraying method (table 1),
Continuously spray 4 weeks (13 days-August 4 days in July).
The medicament and spraying method that the present invention of table 1 uses
The 3rd expansion leaf was taken within 15th respectively with September in 11st, extract RNA, and reverse transcription is c-DNA in July 11, August
Before being sprayed afterwards with fluorescence quantifying PCR method detection, spray relative expression viral in plant leaf afterwards and after the 40d that discontinues medication
Amount, it is specific as shown in Figure 2.
SPVD real-time fluorescence quantitative PCRs primer is synthesized by Shanghai Sheng Gong bioengineering Co., Ltd.
SPFMV primers are：
FSPFMV-3Q:GACTGATATGAGTCTTGCGCGRTATGCG(SEQ ID NO.5)
RSPFMV-2Q:TGTGCCTCTCCGTATCYTCTTCTTGCGT(SEQ ID NO.6)
Objective gene sequence is 149bp.
SPCSV primers are：
FSPCSV-1Q:CCCAACGTGTTTATCTATTACTAAGAGTGG(SEQ ID NO.7)；
RSPCSV-1Q:AATACTGGGGAGCTATCTTACGTTTGA(SEQ ID NO.8)
Objective gene sequence is 170bp.
Treatment group and control group are subjected to real-time fluorescence quantitative PCR detection.
Above-mentioned response procedures are：95℃10min；95 DEG C of 15s, 60 DEG C of 15s, 72 DEG C of 20s, 40 circulations, from 60 DEG C~95
DEG C measure primer melting curve, be with sweet potato IbARF (ADP-ribosylation factor, ADP ribosylation factor) gene
Internal reference, reference gene IbARF primer sequences are
IbARF-L:CTTTGCCAAGAAGGAGATGC(SEQ ID NO.9)
IbARF-R:TCTTGTCCTGACCACCAACA(SEQ ID NO.10)
Objective gene sequence is 185bp.
The relative amount of virus is calculated using 2- Δ Δ Ct methods.Using the initial viral level of each processing sample as 1, each place
Expression quantity of the reason per sub-sampling and the CK of same sample time is contrasted.As a result show, different agents are acted on FMV and CSV
Effect is different.
, will be effective to FMV and two kinds of viruses of CSV according to relative amount viral in treatment group and control group blade
Medicament is divided into four classes：
First kind medicament：No. 7 Physcions (0.1%).FMV and CSV relative amounts in rear Sweet Potato Leaf are sprayed to decline,
Its content does not rise after stopping sprays.This group of medicament can effectively reduce FMV and CSV virus relative amounts, and drug effect is grown.
Second class medicament：No. 2 BASF virus stock solution useds.Rear Sweet Potato Leaf two kinds of viruses of FMV and CSV are relative after spraying medicament
Content declines, and stops spraying rear 40d viral levels as 30% or so of control.This group of medicament can preferably reduce virus and contain relatively
Amount, and drug effect is longer.
3rd class medicament：No. 1 fatty acid mixed (10%), No. 4 Ningnanmycins (8%), No. 13 alkene hydroxyl moroxydines.Spray
Rear Sweet Potato Leaf virus relative amount to be applied to decline, stopping sprays rear 40d viral levels less than control, but more than the 50% of control.
This group of medicament can preferably reduce viral relative amount, but drug effect is shorter.
4th class medicament：No. 5 people's letter-anti-virus, No. 6 viral No. II, No. 10 special pulvis of Garrick capsicum difficult to understand.After just spraying
Two kinds of viral relative amounts decline, but 40d restrovirus contents rise and exceed control group.This group of medicament can reduce disease to a certain degree
Malicious relative amount, but drug effect is short.
According to above-mentioned evaluation index, 0.1% Physcion can effectively reduce the content of virus, and sweet potato SPVD is prevented and treated and imitated
Fruit is optimal.
3. influence of the chemicals treatment to sweet potato SPVD Plant agronomic traits
In September 15 days, it is total to measure the most long climing length of each treatment group, chlorophyll content (SPAD-502 chlorophyll meters), underground part
Fresh weight, overground part fresh weight, total biomass.
After chemicals treatment, as shown in figure 3, except the chlorophyll content of No. 8 and No. 12 treatment group sweet potatoes is relatively compareed without significantly change
Change.The chlorophyll content of other treatment group sweet potatoes is significantly or pole is significantly higher than control.
Different agents are also different to the action effect of the climing length of sweet potato, as shown in figure 4, No. 5, No. 10, No. 12, No. 14 4 kinds of medicines
Agent does not make significant difference to climing length, and the climing length of other treatment group sweet potatoes is significantly or pole is significantly higher than control.
After medicament, as shown in figure 5, in addition to No. 5 medicaments do not make significant difference to the underground part fresh weight of sweet potato, other medicaments
The underground part fresh weight of sweet potato has compared with control and significantly or extremely significantly increased after processing.
After medicament is applied, as shown in fig. 6, in addition to No. 1, No. 5, No. 10 3 kinds of medicaments, other chemicals treatment group overground parts are fresh
Weight is notable or pole is significantly higher than control.
As shown in fig. 7, sweet potato total biomass is influenceed using medicament it is also larger, No. 5, No. 10, it is right after No. 14 this 3 kinds of medicaments
The total biomass of sweet potato is not made significant difference, and the total biomass of other chemicals treatment group sweet potatoes has compared with control significantly or extremely significantly to be increased
It is long.
4. the overall merit of pharmacy effect effect
By relative amount overall merit viral in every physical signs of sweet potato after chemicals treatment and sweet potato, screening pair
The preferable medicament of SPVD prevention effects.
After chemicals treatment, the indexs such as physical signs and the economical character of sweet potato are made to have notable or pole to significantly improve compared with control
Medicament have No. 2, No. 3, No. 4, No. 6, No. 7, No. 9, No. 11, No. 13, totally 8 kinds (table 2).After chemicals treatment, in sweet potato viruses seedling
Declining larger medicament with respect to viral level has No. 2, No. 7.
The overall merit of the chemicals treatment restrovirus content of table 2 and economical character
In summary index, 0.1% Physcion can not only effectively reduce relative amount viral in sweet potato, moreover it is possible to aobvious
Write and improve sweet potato items physical signs, greatly reduce production loss caused by SPVD.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modification, equivalent substitution or improvement made within refreshing and principle etc., should be included in the scope of the protection.
SEQUENCE LISTING
<110>Xuzhou Agriculture Science Inst., Xuhuai area, Jiangsu（Xuzhou Sweetpotato Center）
<120>A kind of method for mitigating sweet potato SPVD harm
<160> 10
<170> PatentIn Version 2.1
<210> 1
<211> 34
<212> DNA
<213>Artificial sequence
<400> 1
CCGGGTCTRRTGAGARHACTGAATTTAAAGATGC 34
<210> 2
<211> 32
<212> DNA
<213>Artificial sequence
<400> 2
GACTCTCGAGCCTATTGCACACCCCTCATTCC 32
<210> 3
<211> 22
<212> DNA
<213>Artificial sequence
<400> 3
GACGGKGGTACKATGAARGTCC 22
<210> 4
<211> 26
<212> DNA
<213>Artificial sequence
<400> 4
GGCTCACAAACHGAYTTCATAAACAT 26
<210> 5
<211> 28
<212> DNA
<213>Artificial sequence
<400> 5
GACTGATATGAGTCTTGCGCGRTATGCG 28
<210> 6
<211> 28
<212> DNA
<213>Artificial sequence
<400> 6
TGTGCCTCTCCGTATCYTCTTCTTGCGT 28
<210> 7
<211> 30
<212> DNA
<213>Artificial sequence
<400> 7
CCCAACGTGTTTATCTATTACTAAGAGTGG 30
<210> 8
<211> 27
<212> DNA
<213>Artificial sequence
<400> 8
AATACTGGGGAGCTATCTTACGTTTGA 27
<210> 9
<211> 20
<212> DNA
<213>Artificial sequence
<400> 9
CTTTGCCAAGAAGGAGATGC 20
<210> 10
<211> 20
<212> DNA
<213>Artificial sequence
<400> 10
TCTTGTCCTGACCACCAACA 20
Claims (3)
- A kind of 1. method for mitigating sweet potato SPVD harm, it is characterised in that use concentration to be made into for 0.1% Physcion 300 times of dilutions, carry out dispenser by the way of to sweet potato foliar spray.
- 2. a kind of method for mitigating sweet potato SPVD harm according to claim 1, it is characterised in that use Physcion Dilution, spray Sweet Potato Leaf weekly after sweet potato growing 1 time, continuously spray 4 weeks.
- 3. a kind of evaluation method of the sweet potato SPVD extents of injury, it is characterised in that power is sprayed by fluorescence quantifying PCR method measure Profit requires two kinds of viral levels of FMV and CSV in 1 or 2 medicament rear blades, determines sweet potato physical signs and economical character, with evaluation Mitigation effect of the medicament to sweet potato SPVD.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110199881A (en) * | 2019-07-10 | 2019-09-06 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of sweet potato health seedling cultural method removing sweet potato viruses complex SPVD |
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| CN103849691A (en) * | 2014-02-26 | 2014-06-11 | 福建省农业科学院作物研究所 | Sweet potato virus detection primers and method |
| CN103937908A (en) * | 2014-03-28 | 2014-07-23 | 河南省农业科学院植物保护研究所 | Real-time fluorescence quantitative PCR detection method of West African strain of sweet potato chlorotic stunt virus and application |
| CN104430349A (en) * | 2014-12-22 | 2015-03-25 | 成都新朝阳作物科学有限公司 | Pesticide composition capable of preventing and treating plant virus diseases and application of pesticide composition |
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| Title |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110199881A (en) * | 2019-07-10 | 2019-09-06 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of sweet potato health seedling cultural method removing sweet potato viruses complex SPVD |
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Application publication date: 20171121 |